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ISSN 1313 - 8820Volume 7, Number 1

March 2015

2015

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Genetics and Breeding

Atanas Atanasov (Bulgaria)Nikolay Tsenov (Bulgaria)Max Rothschild (USA)Ihsan Soysal (Turkey)Horia Grosu (Romania)Bojin Bojinov (Bulgaria)Stoicho Metodiev (Bulgaria)

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Nikolai Todorov (Bulgaria)Peter Surai (UK)Zervas Georgios (Greece)Ivan Varlyakov (Bulgaria)

Production Systems

Dimitar Pavlov (Bulgaria)Bogdan Szostak (Poland)Dimitar Panaiotov (Bulgaria)Banko Banev (Bulgaria)Georgy Zhelyazkov (Bulgaria)

Agriculture and Environment

Georgi Petkov (Bulgaria)Ramesh Kanwar (USA)Martin Banov (Bulgaria)

Product Quality and Safety

Marin Kabakchiev (Bulgaria)Stefan Denev (Bulgaria)Vasil Atanasov (Bulgaria)

English Editor

Yanka Ivanova (Bulgaria)

2015

ISSN 1313 - 8820 Volume 7, Number 1March 2015

Effect of black (stem) rust (Puccinia Graminis F.SP. Tritici) attack to the spike characteristics in Polish wheat (Triticum Polonicum L.)

H. Stoyanov*

Plant Growing Department, Faculty of Marine Sciences and Ecology, Technical University, 1 Studentska, 9000 Varna, Bulgaria

Abstract. The obtaining of high yields of crops is directly dependent on the cultivated varieties. In their creation it is essential the selected initial breeding material to possess certain qualities that lead to overcoming the effects of biotic and abiotic stress factors. Common winter wheat is characterized by great diversity of possibilities for combining initial breeding material due to phylogenetic similarity to many species of the genera Triticum, Aegilops, Secale. In creation of synthetic hexaploids (2n = 6x = 42, AABBDD), the choice of tetraploid component involved into the crosses is very important. The species Triticum polonicum (2n = 4x = 28, AABB) possesses many valuable features related to its protein content, resistance to brown rust, powdery mildew and septoria leaf blight, but it is susceptible to varying levels of black rust attack. This is a prerequisite a correlation between the attack of the pathogen and some spike indicators that correlate with grain yield to be searched. To determine the relationship between the attack and the specific parameters, 52 accessions of the species Triticum polonicum have been studied in the financial 2012/2013 year, to the following indicators: length of spike, length of spike with awns, number of spikelets, weight of spike, weight of grains per spike, number of grains per spike, weight of 1000 grains, weight of spike after threshing. An evaluation of the infectious type of black rust attack on each accession is recorded according to 9-point scale, and correlation with the average values of each of the spike indicators is reported. To neutralize the influence of the factor 'accession' two specific indexes are calculated as a corrected value of the weight of 1000 grains. The highest and significant correlation was observed between the infectious type and both specific index (-43.2% and -44.6%), and less with the weight of 1000 grains (-41.9%). Insignificant or unreliable is the correlation of the other spike indicators with the infectious type. The lack of reliable and significant correlation of pathogen attack with the weight of grains per spike as an essential element of the yield, gives rise to claims that Triticum polonicum possesses a compensatory mechanisms to the effect of the black rust. This determines most of the studied accessions as a potential initial material for the creation of synthetic wheat or for inclusion into the selection programs of winter wheat.

Keywords: black rust, spike morphology, polish wheat

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 7, No 1, pp , 201525 - 35

Introduction (Zhang et al., 2008; Spetsov et al., 2008; Spetsov et al., 2009; Stoyanov et al., 2010). This makes them a valuable source for transfer of biotic and abiotic stress factors resistance genes. The bread wheat demand in the world is steadily increasing due Depending on the tetraploid component involved in the output to population growth, especially in developing countries. This leads crosses, hexaploid synthetic wheat exhibits different properties with to the development of new production technologies, and to the regards to its resistance to diseases. Regardless of the involved wild creation of new wheat varieties which are to meet the increasingly diploid component accession (Aegilops tauschii (2n = 2x = 14, DD), extensive characteristics of agricultural production. By classical with high resistance to pathogens of powdery mildew, brown rust, breeding, varieties that combine high productivity, high quality septoria leaf blight and black rust are distinguished synthetic lines parameters (protein and amino acid composition), resistance to involving Triticum dicoccoides, Triticum dicoccon, Triticum durum biotic and abiotic stress factors (resistance to economically (Spetsov et al., 2009; Stoyanov, 2012a, Zhang et al., 2008). Valuable important diseases - powdery mildew, brown rust, septoria leaf source as an initial component for creating synthetic hexaploid blight, fusarium head blight, black rust, resistance to lodging, wheat has the species Triticum polonicum (Kang et al., 2008; tolerance to drought, reduced susceptability to metal toxicity) are Dorofeev, 1987).created (Stoyanov et al., 2010; Stoyanov, 2012a; Stoyanov, 2013).

Polish wheat Triticum polonicum (2n = 4x = 28, AABB), belongs Combining a number of important parameters of plants requires to the group of tetraploid naked wheats (Wiwart et al., 2013). Its thorough selection of initial breeding material, which possesses a occurrence as crop is low as it is sown in small areas mainly in particular characteristics, but also much of hybridisation work. Southern Spain (Andalusia), Italy, Western Ukraine, Algeria and Therefore, the information of the accessions involved in the crosses Ethiopia, and in the warm regions of Asia (Wiwart et al., 2013; became the basis for quick and high-quality breeding work.Rodriguez-Quijano et al., 2003; Eticha et al., 2006). The species is As a primary tool of the classical breeding, wide hybridization distinguished by its specific spike morphology - highly elongated features a number of difficulties such as weak crossability, low glumes reaching 4 cm (Wang et al., 2002; Wiwart et al., 2013). The viability, high sterility of the obtained plants (Stoyanov, 2013; grains are not easily released of the spikes, and it is characterized by Naskidashvili, 1984). However, it is reported for many created its size and weight (M1000 up to 80g.), high protein content. Despite hybrids and amphidiploids into tribus Triticae, which are important their negative qualities such as low tillering, prone to lodging and a for the bread wheat breeding (Stoyanov, 2013). Particularly height of stem, the species is characterized by high resistance to important are hexaploid synthetic wheat lines (2n = 6x = 42, some fungal pathogens such as powdery mildew, brown rust and AABBDD), which possess identical genomic constitution with septoria leaf blight, but also by a strong variation to the attack of common winter wheat Triticum aestivum (2n = 6x = 42, AABBDD)

25

* e-mail: [email protected]

26

black rust (Wiwart et al., 2013; Eticha et al., 2006; Yacubtsiner and 1000 grains – a specific index eliminating the factor influence (SEFI) Shopina, 1970). This implies the presence of resistance genes in – ratio between M1000 and the difference between WS and WGS; current accessions of the species that could be significant in the SEFI2 – ratio between M1000 to the product of the difference creation of synthetic hexaploids (Wiwart et al., 2013; Rodriguez- between WS and WGS and NSS. The corrective parameters are Quijano et al., 2003). Black rust is a highly damaging disease determined based on the hypothesis that black rust does not affect causing serious reduction in the yield of different wheat species the number of the formed spikelets and the weight of the vegetative (Haile et al., 2012). The symptoms are associated with damage to all part of the spike as it is an obligate pathogen as Beteselassie et al. aboveground plant organs, primarily the stem, and in strong attack it (2007a) stated. This allows avoiding differences caused by genetic reaches the spike attacking the glumes, lemmas and paleas (Roelfs characteristics of each accessions.et al., 1992). The presence of large glumes in Polish wheat, which is The data is averaged (AV) and a standard deviation (SD) is associated with an increase in the photosynthetic activity of the reported, per accessions and in general. The total variation plants (Wang et al., 2002), creates specific conditions for more coefficient (VC) and the statistical error per accessions (SE) are vigorous development of the pathogen onto the spike. Greater calculated. A correlation analysis was made between all properties distribution near the reproductive organs suggests a significant and infectious type of black rust. The significance of obtained results decrease in the value of seedset into the infested spikes (Roels et is reported. To summarize the data and for analysis of variance al., 1992; Stoyanov, 2012b). Thus, the disease leads to changes in software Microsoft Excel 2003 was used and for correlation analysis the morphological characteristics of the spikes, which makes – IBM SPSS Statistics 19. Total, direct and indirect effects of ITP to possible to establish a correlation between the infection type, which other properties are established by IBM SPSS Amos 20.characterizes the resistance level and the properties of the spike Evaluation of the suitability of the accessions as initial breeding related to the productivity of the accessions (Roelfs, 1988). material is made on the basis of a 3-point scale of the pathogen

The purpose of this report is to determine the relation between reaction of black rust and also to the compensatory capabilities of infectious type under attack of black rust and spike characteristics of the accessions (Stoyanov, 2012a).Polish wheat under natural infection background and to make an evaluation of the suitability of species to participate as an initial material into the creation of synthetic hexaploid wheats. Results and discussion

The results of morphological and physiological characteristics Material and methods of the Polish wheat accessions are presented in Tables 1, 2, 3 and 4.

From the data it could be traced a wide variation in some of the Fifty two accessions of Polish wheat Triticum polonicum, morphological characteristics of the spikes. Particularly high levels

originating from the collection of IPK-Gatersleben-Germany were of variation are observed in the indicators NSLS, WDLS and specific used. 15 seeds of each accession were sown with a row spacing 30 indices SEFI and SEFI2. In the first two indicators results are due to cm and 5 cm inside rows. Sowing was carried out on 03.05.2013, the uneven distribution of spikelets in different accessions, which under field conditions in the area of Stozher, Dobrich region. sets very thin or very thick spikes. In the second two indexes the

To determine the infectious type of black rust pathogen, ten greater variation is due to the neutralization of the influence of the point scale, aligned to the appropriate scale for susceptibility to factor 'accession', which leads to the protrusion of the reaction of the brown rust of Cereal Rust Laboratory, Minnesota, USA was used pathogen attack of black rust and uneven seedset in individual (Stoyanov, 2014a). Evaluation has been conducted under field spikelet.conditions in 2013 in phase lactic maturity of accessions by naturally The effect of the attack by the pathogen to the number of presented races of the pathogen. For proper identification of the spikelets per spike (Table 1) could be regarded as a insignificant due pathogen and reporting of its appearance two highly susceptible to the mechanism of pathogenicity impact. The disease primarily accessions of species Triticum vavilovii are used, which are highly affects the stem and secondary glumes and partly lemmas of attacked by the pathogen of black rust according to Stoyanov already formed spikes (Roefls et al., 1992; Stoyanov, 2012b). During (2012b). the attack, previous inoculation and development of the disease

Harvesting is done in phase full maturity between 12 and does not coincide with the period of the formation of spikes spindle 20.07.2013. Of each sample 10 fully matured spikes were randomly (Stoyanov, 2012b). Although the studied accessions are spring selected with established or not established presence of black rust forms, the black rust infection occurs in the period after 30.05 when damage, but not attacked by other pests. Morphological evaluation accessions are in a period of active stem extension and spikelets are of spikes of each accession is done on 6 quantitative: length of spike already formed, and some of the plants have reached stage 'flag (LS), length of spike with awns (LSA), weight of spike (WS), number leaf'. Despite the established correlation of over 30%, it is unreliable of spikelets per spike (NSS), weight of grains in a spike (WGS), and insignificant.number of grains in a spike (NGS) and 6 index properties: awness The length of spike (Table 1) is a too variable indicator which is index (AI) – ratio of LSA and LS, the weight distribution along the formed under the influence of environmental factors mainly related length of spike (WDLS) – ratio of WS and LS, number of spikelets to to the rate of growth and development – the quantity of available the length of spike (NSLS) – ratio of NSS and LS, average weight of nutrients in the soil, atmospheric and soil moisture, temperature, spikelet (WSS) – ratio of WS and NSS, weight of 1000 grains pathogens influence (Rachovska and Uhr, 2010; Nikolova and (M1000) – set according to standard methodology of BDS, Panayotov, 2008). However, the limit of LS was determined by percentage of grain weight in a spike (GI) – ratio between WGS and hereditary factors (Stoyanov, 2014b). As accessions grown under WS. the same conditions of fertilization and climate, variability in

To neutralize the influence of the factor "accession" two specific characteristics probably is due to the influence of pathogens and weight indexes are calculated, as the corrected value of the weight of heredity. Spikes actively grow from phase 'flag leaf' to beginning of

27

Table 1. Morphological characteristics of studied Triticum polonicum accessions by properties NSS, LS, LSA and WS

NSS

AV AV AV AVSD SD SD SDSE SE SE SE

LS LSA WSAccessions

TRI642TRI768TRI889TRI891TRI893TRI933TRI1896TRI1897TRI1950TRI1951TRI2223TRI2997TRI3203TRI3248TRI3289TRI3311TRI3408TRI3410TRI3412TRI3428TRI3440TRI3441TRI3442TRI3478TRI3550TRI3597TRI4090TRI4342TRI4344TRI5358TRI5915TRI6961TRI6962TRI9650TRI9924TRI15114TRI17827TRI19040TRI19053TRI19111TRI19114TRI19168TRI19195TRI19196TRI19211TRI19226TRI19230TRI19244TRI28850TRI28851TRI28852TRI28855TotalVC, %

14.2015.8012.5015.5013.2013.9015.4015.3013.3014.1012.7015.6016.8014.8015.3013.3015.2015.2014.8014.6015.5614.2015.9016.1014.2016.8011.5014.7012.6012.5613.8017.1015.1014.0013.6013.5013.7014.5013.8015.3010.1117.2013.0014.5012.0014.2015.7013.0014.2017.6016.0018.1014.5716.03

3.031.481.721.901.811.372.321.701.341.521.832.121.481.553.201.251.481.402.251.352.602.781.791.520.631.322.351.701.262.192.201.372.021.631.651.270.951.721.991.772.091.691.943.341.891.551.341.891.872.071.151.732.34

1.360.470.540.600.570.430.730.540.420.480.580.670.470.491.010.400.470.440.710.430.870.880.570.480.200.420.960.540.400.730.700.430.640.520.520.400.300.540.630.560.700.530.611.060.600.490.420.600.590.650.370.550.10

74.6089.5072.7093.50

102.00116.1098.8085.00

105.90116.4092.1055.8099.6094.8064.7087.2083.6081.6098.7071.30

105.8991.30

102.0087.10

109.40111.5068.1790.30

107.1090.1168.4082.5074.80

111.7067.1072.3068.4076.2079.20

101.4074.2291.9053.6069.2063.4077.6082.1087.8092.50

108.9075.4069.5086.6823.13

16.7931.7512.1411.3714.7311.2511.3210.5514.0011.9918.488.389.92

10.4215.047.817.90

10.3317.755.77

23.9219.1713.386.08

10.0515.5914.5215.3410.2210.6510.326.008.89

12.247.816.274.488.13

11.4113.7716.2215.645.95

16.0510.5210.6410.8513.0211.9010.688.189.03

20.05

7.5110.043.843.594.663.563.583.344.433.795.852.653.143.304.762.472.503.275.611.837.976.064.231.923.184.935.934.853.233.553.261.902.813.872.471.981.422.573.614.355.414.951.885.073.333.363.434.123.763.382.592.860.89

201.80161.80133.40159.60188.50204.20174.30165.50197.10208.90182.20118.00166.90181.60153.90165.20179.10179.40184.50186.60161.11161.60171.60188.60189.20176.00132.50162.90203.00170.78184.5082.5074.80

205.20171.30188.80163.10175.20172.40187.90163.22154.20167.90183.40176.60178.10184.20160.20153.90171.80180.90127.10169.5617.96

19.1214.0514.6113.7824.4110.3622.7815.0215.9419.4423.3311.7913.1614.0317.5011.6810.0915.7019.3410.7025.0120.4718.127.71

28.2525.8215.3221.2415.5715.3510.886.008.89

15.2311.289.16

13.3010.7317.5411.3824.4421.0614.3215.7110.9314.5515.0817.6219.1319.549.69

12.2630.45

8.554.444.624.367.723.287.214.755.046.157.383.734.164.445.533.693.194.966.123.388.346.475.732.448.938.166.256.724.925.123.441.902.814.823.572.904.213.395.553.608.156.664.534.973.464.604.775.576.056.183.063.881.35

3.612.491.612.662.442.942.873.202.723.091.951.432.973.292.461.783.282.992.153.822.822.162.663.442.763.531.592.402.892.463.663.182.363.243.392.972.372.932.342.671.813.482.443.062.792.993.261.852.783.264.693.082.79

32.95

1.240.900.650.700.650.480.940.590.690.650.920.670.780.441.050.320.540.650.620.631.410.750.860.690.260.530.800.830.290.780.960.520.650.570.800.470.390.650.600.560.920.800.520.951.000.890.900.560.590.510.600,620.92

0.560.290.210.220.200.150.300.190.220.210.290.210.250.140.330.100.170.200.200.200.470.240.270.220.080.170.330.260.090.260.300.160.210.180.250.150.120.210.190.180.310.250.160.300.320.280.280.180.190.160.190.200.04

28

flowering and their growth ends after seed formation. In some combination of the formation of a smaller number of grains and accessions this period coincides with the period of black rust therefore a reduction of the absolute weight of the grains as a result development. As spike spindle is not attacked by the pathogen, of suppressed growth of infected plant, particularly during grain independently of the phase and of infection, its growth could be formation (Roefls, 1988). Since the grains are not directly attacked influenced only indirectly, by inhibiting its growth. Obligate type of by the pathogen, the impact to WGS (Table 2) is more indirect than development of black rust does not allow a strong suppression of direct. Nutrients that determine growth and grain filling in heavy plant growth (Beteselassie et al., 2007a). In many cases it is infestations, are limited to the development of the fungus. As black reported that the biomass of the plant does not vary widely in rust is an obligate parasite, the fungus seeks to maintain maximum infested and not infested plants (Roefls et al., 1992; Cox et al., 1987; plant live a long time. Therefore, a certain part of the nutrients are Singh et al., 1998). As the primary attack is on the stems of plants consumed by the plant, in order to maintain its vegetative mass. and photosynthetic activity is not reduced in a wide range, it is likely However, due to shortage of nutrients the formed grain remains length of spike to be affected to a minor extent, exept at high attack. undeveloped. This is part of the pathogen strategy (Ayala and Kiger, Accessions TRI1896, TRI3440 and TRI3442 are with length of spike 1987), as consequently from the small grains are formed weaker above the average of accessions. This together with unreliable and plants which are more susceptible to attack during the next growing insignificant correlation of ITP to LS, highlights the weak impact of season (Roelfs et al., 1992). In this case, although the influence is the attack of black rust (Table 5). indirect as a mechanism, the effect on the index is direct. This

Awns of cereals are genetically determined property and their defines the identical values of the correlation coefficient and the formation and development coincides with that of the spike spindle direct effect of the infectious type to WGS (Table 5 and 6).(Elbaum et al., 2007). The presence of fewer stomata and typical Number of grains in the spike (Table 2) in certain spikes of the awn design prevents normal process of inoculation. They are examined accessions is highly lowered, or even seeds are absent or located at a certain height and due to the small surface moisture underdeveloped after fertilization and embryo formation. As glumes droplets do not retain long enough. This prevents the germination of are actively attacked by black rust, the spikelets under the influence the spores as they need condensed water (Roefls et al., 1992). Black of the mycelium of the fungus is highly distorted, leading to forced rust does not attack the awns of the studied Polish wheat crushing of lemmas and paleas (Figure 1 A,B). Since the integrity of accessions. This is probably due to elongated glumes which the florets is a prerequisite for the proper functioning of the formation of plants possess. This creates enough nutrient area for the the grain (Ganusheva et al., 2013), the damaged lemmas do not development of the pathogen and its passage on awn worn by allow embryo development and endosperm formation. Therefore, lemmas is delayed. When a sufficient amount of inoculum is the effect of the pathogen can again be considered to be indirect, presented, the attack is limited because at this stage awns are in the since it does not directly influence the reproductive apparatus of the beginning of yellowing and they dies. In some susceptible plants. In the accessions that are less attacked by the pathogen, accessions from other wheat species, strong attack on awns is such distortion of the spikes is not obtained (Figure 1 D,E,F), and the reported, but after flowering phase (Stoyanov, 2012b). A similar type grain is formed, but it has a reduction in the absolute weight. The of attack does not imply severely limiting their growth, as it ends direct effect on this indicator again coincides with the correlation with before flowering of the spikes. For this reason, the attack of the black ITP, which highlights the insignificance of the impact of the disease rust pathogen does not affect LSA (Table 1). This is evidenced by the on the number of grains in the spike (Tables 5 and 6).lack of significant and reliable correlation of the indicator with ITP The awness index (Table 2) as a complex indicator shows (Table 5) and by the unawned accession with low pathogenic attack unreliable correlation with ITP. This is due to the fact that its (Figure 1C). components LS and LSA are not directly affected by the disease,

Weight of spike (Table 1) is a complex indicator that includes a and are formed by the action of other factors. Therefore, the weight of grains that are formed in the spike and the dry biomass of presence and development stage of the awns is not affected by the spike spindle, glumes, lemmas, paleas and awns. Black rust affects pathogen, but rather than hereditary factors.plants productivity by reducing their yield decreasing the weight of Dry biomass of the spikes expressed by the indicator PSS single grains or preventing the grain formation in separate florets, (Table 2) allows to quantify the vegetative part of the spikes. Also like spikelets or even the entire spike (Roefls et al., 1992; Roefls, 1988; the indicator LS, this indicator and its formation is directly dependent Stoyanov, 2012b). Therefore, it is impossible to report the direct on hereditary factors that determine the limits, but also is under the impact of the attack of pathogen on this indicator because it is influence of quantitative and qualitative contents of nutrients and difficult to determine on which, and in what degree of its components moisture in the soil. For this reason and due to the fact that the black are influenced by disease. However, some effects could be rust is an obligate pathogen, it would not exhibit a substantial direct determined indirectly, as strong attack inhibits growth (Pfender, effect on the morphology of the spikes. Indirect effect related to the 2004). This leads to poor development of additional tillers, especially amount of biomass would occur at high attack and suppresses the in early attacks (Pfender, 2004). They form spikes, but they are growth of additional tillers, like WS indicator (Stoyanov, 2012b). In lighter, due to the small number of fertile florets and form smaller and this case, a similar effect could be sought as a correlative stunted or shriveled grains. Since the main tiller is less affected, this relationship between the standard deviation, which shows the suggests heterogeneity of the weight of spikes in one plant. The greater variation and infectious type. The spikes in the accessions more different are the spikes, the greater is the variation, and which are attacked to a greater extent, in fact, exhibit a great therefore the standard deviation. Accessions as TRI1896, TRI3440 variability in this parameter. This highlights the indirect effect on the and TRI3442 exhibit high values o f standard deviation of WS, which indicator, but the lack of correlation with average excludes the direct coincides with their high infectious type of damage. impact (Table 5).

Reducing the weight of the grains in bread wheat under attack Due to the insignificant impact that black rust has on NSS and of black rust was reported by a number of authors and researches LS, as components of the indicator NSLS (Table 3) (Pinthus and (Roelfs et al., 1992; Villareal et al., 1991; Singh et al., 2006; Millet, 1978), it can be assumed that under the attack of the Stoyanov 2012b; Dochev et al., 2009). Reduced productivity is pathogen the influence will not be so significant. Insignificant and

Table 2. Morphological characteristics of studied Triticum polonicum accessions by properties WGS, NGS, AI and PSS

WGS


NGS AI PSSAccessions


2.331.571.021.831.672.111.722.101.942.241.460.811.972.271.651.242.292.171.532.781.501.281.542.361.922.051.031.372.031.852.712.431.692.302.542.201.672.161.681.891.302.551.822.112.021.962.241.272.012.273.442.231.93

37.18

0.660.800.570.560.470.250.970.320.520.440.560.620.630.330.810.210.400.440.440.450.980.400.780.630.280.530.790.770.160.600.740.410.510.350.570.370.290.470.430.340.640.600.380.700.680.780.750.440.440.600.430.480.72

0.300.250.180.180.150.080.310.100.170.140.180.200.200.100.250.070.120.140.140.140.330.130.250.200.090.170.320.240.050.200.230.130.160.110.180.120.090.150.140.110.210.190.120.220.210.250.240.140.140.190.140.150.03

34.6036.3025.9037.8026.9032.3032.7040.2028.2033.2028.1026.4037.2033.4032.1024.8034.4035.8028.5036.2031.4430.0037.6038.6030.6040.2022.3328.2029.8027.0033.7046.8030.6030.5032.0030.6027.5035.1026.7033.7017.7844.7031.5033.4026.6033.7038.6026.6039.7047.8043.9047.2033.1830.35

10.3613.219.567.876.142.26

18.975.516.615.378.25

15.028.164.65

13.743.585.345.206.805.77

15.735.799.838.033.756.20

14.9212.222.867.257.945.737.854.905.874.625.257.556.456.246.657.895.48

10.338.479.689.317.937.104.106.497.48

10.07

4.634.183.022.491.940.726.001.742.091.702.614.752.581.474.341.131.691.652.151.825.241.833.112.541.191.966.093.860.902.422.511.812.481.551.861.461.662.392.041.972.222.491.733.272.683.062.942.512.251.302.052.370.45

2.783.221.851.721.861.771.771.951.871.802.002.131.681.932.451.902.152.211.902.631.551.801.692.171.741.581.981.821.901.902.741.001.001.842.572.632.392.312.191.872.231.693.172.722.842.322.261.831.671.592.421.842.05

39.69

0.424.880.120.110.180.120.130.080.120.080.160.170.070.160.350.110.140.130.180.170.180.160.080.110.280.130.230.190.170.100.310.000.000.100.190.240.140.180.180.140.150.110.450.380.360.260.200.100.150.190.200.100.81

0.191.540.040.030.060.040.040.030.040.030.050.050.020.050.110.030.040.040.060.050.060.050.020.030.090.040.090.060.050.030.100.000.000.030.060.070.050.060.060.050.050.040.140.120.110.080.060.030.050.060.060,030.04

1.280.920.590.830.770.841.161.100.770.860.490.620.991.020.810.540.980.830.631.041.310.881.121.080.831.480.571.030.860.620.950.750.670.940,850.770.700.770.660.790.500.930.620.940.771.031.020.580.770.991.250.840.86

39.85

0.630.220.200.190.220.260.450.300.180.300.460.130.270.240.310.120.230.260.190.230.700.420.290.290.140.400.250.150.180.210.240.180.210.240.260.140.140.210.180.240.290.220.150.370.350.250.300.140.220.640.240.210.34

0.280.070.060.060.070.080.140.100.060.100.140.040.090.080.100.040.070.080.060.070.230.130.090.090.040.130.100.050.060.070.080.060.070.080.080.040.040.070.060.070.100.070.050.120.110.080.100.040.070.200.080.070.02

29

30

Table 3. Morphological characteristics of studied Triticum polonicum accessions by properties NSLS, WDLS, WSS, M1000

NSLS


WDLS WSS M1000Accessions


0.190.330.170.170.130.120.160.180.130.120.140.280.170.160.240.150.180.190.150.210.150.160.160.190.130.150.170.170.120.140.200.210.200.130.200.190.200.190.180.150.140.190.240.210.190.180.190.150.150.160.210.260.18

48.21

0.010.550.010.010.020.010.020.030.010.010.020.020.010.010.040.020.010.010.010.020.030.020.020.020.010.020.010.030.010.010.010.020.020.010.020.010.010.010.020.010.010.030.020.020.010.010.010.010.010.010.020.020.09

0.000.170.000.000.000.000.010.010.000.000.010.010.000.000.010.000.000.000.000.010.010.010.010.000.000.010.000.010.000.000.000.010.010.000.010.000.000.000.010.000.000.010.010.000.000.000.000.000.000.000.010.010.00

0.050.050.020.030.020.030.030.040.030.030.020.020.030.030.040.020.040.040.020.050.030.020.030.040.030.030.020.030.030.030.050.040.030.030.050.040.030.040.030.030.020.040.050.040.040.040.040.020.030.030.060.040.03

48.69

0.010.090.010.000.000.000.010.000.000.000.010.010.010.000.010.000.000.000.000.010.010.000.010.010.000.000.010.010.000.010.010.000.010.000.010.000.000.010.010.000.010.000.010.010.010.010.010.000.000.000.010.000.02

0.000.030.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.00

0.250.150.120.170.180.210.190.210.200.220.150.090.180.220.160.130.220.200.140.260.180.150.170.210.190.210.130.160.230.190.260.190.150.230.250.220.170.200.170.170.170.200.190.210.230.210.210.140.200.190.290.170.19

27.35

0.040.050.040.030.040.020.060.040.040.030.050.040.040.020.050.020.030.030.030.030.070.040.050.030.020.030.040.050.020.030.040.020.030.020.040.020.020.030.030.020.050.030.030.060.050.060.050.030.030.030.030.020.05

0.020.020.010.010.010.010.020.010.010.010.020.010.010.010.020.010.010.010.010.010.020.010.020.010.010.010.020.020.010.010.010.010.010.010.010.010.010.010.010.010.020.010.010.020.020.020.020.010.010.010.010.010.00

68.2741.2635.7347.9261.7965.1343.0352.2868.4567.2150.9323.4552.6068.0545.1949.9266.7160.2453.0076.8547.2542.4938.7660.5962.7850.0036.8741.9768.3667.7079.5951.8154.8175.4579.0171.8360.9861.4562.7856.1470.4756.5657.5764.8375.5256.3956.3847.1450.6147.0479.1047.3157.2926.92

7.7016.6513.736.447.735.25

24.723.234.055.015.09

15.4213.545.35

16.885.036.485.943.624.21

13.6010.3413.596.713.259.26

19.6719.826.414.625.334.405.882.643.614.003.183.255.403.71

10.494.965.74

14.263.45

13.5813.696.286.699.498.756.84

15.42

3.445.264.342.042.441.667.821.021.281.591.614.884.281.695.341.592.051.881.151.334.533.274.302.121.032.938.036.272.031.541.681.391.860.831.141.271.011.031.711.173.501.571.824.511.094.294.331.982.123.002.772.160.68

Table 4. Morphological characteristics of studied Triticum polonicum accessions by properties GI, SEFI, SEFI2 and ITP

GI


SEFI SEFI2 ITPAccessions


0.660.580.580.680.680.720.530.660.710.730.830.470.650.690.620.700.700.730.710.730.520.600.550.680.700.580.580.510.700.750.740.760.710.710.750.740.700.740.720.710.720.730.750.690.730.630.670.680.730.700.740.730.68

20.40

0.080.200.210.050.040.060.290.040.020.060.380.270.110.060.220.030.050.040.030.030.220.080.170.090.060.140.290.230.040.030.030.040.060.040.030.030.040.030.030.030.030.020.020.070.040.120.140.040.050.190.030.040.14

0.030.060.070.010.010.020.090.010.010.020.120.090.030.020.070.010.020.010.010.010.070.030.050.030.020.040.120.070.010.010.010.010.020.010.010.010.010.010.010.010.010.010.010.020.010.040.040.010.010.060.010.010.01

73.4146.2370.2459.4285.7691.8444.1550.2193.3086.0575.3838.7856.9669.9361.5795.9770.3777.7390.3677.4346.5654.7337.6259.2377.2837.9085.4040.3682.64

119.3988.3672.1690.0785.9899.3095.6990.3084.93

102.7576.12

168.6962.7396.7080.09

117.1457.8959.4683.2572.2126.8965.0459.2674.5449.10

55.4121.5848.8011.5826.6958.2532.3111.5824.8325.1262.2825.7022.3516.0931.9320.8012.9216.9322.5718.3329.0023.2817.1615.3412.4916.8157.4519.8220.1634.4121.3416.9532.6225.8422.1417.4522.0122.9733.2619.9958.1110.9219.8234.8749.8522.9222.1614.1027.4549.2211.1615.3136.60

24.786.82

15.433.668.44

18.4210.223.667.857.94

19.698.137.075.09

10.106.584.085.357.145.809.677.365.434.853.955.32

23.456.276.37

11.476.755.36

10.328.177.005.526.967.27

10.526.32

19.373.456.27

11.0315.767.257.014.468.68

15.563.534.841.62

6.152.955.653.946.807.013.003.307.286.296.052.443.414.834.007.314.705.216.445.433.114.082.463.735.462.318.202.726.76

10.036.714.276.246.417.557.226.696.087.995.13

18.323.737.776.10

10.604.223.826.635.331.594.093.355.54

69.64

6.171.504.041.122.895.762.590.783.062.325.631.701.371.482.301.871.141.422.641.842.172.031.321.110.981.127.431.422.454.042.481.123.112.752.492.032.032.344.411.799.370.992.603.256.222.301.481.872.612.960.841.063.86

2.760.481.280.350.911.820.820.250.970.731.780.540.430.470.730.590.360.450.830.580.720.640.420.350.310.363.030.450.781.350.790.360.980.870.790.640.640.741.400.563.120.310.821.031.970.730.470.590.830.940.270.340.17

8.009.006.008.005.007.009.008.006.006.007.008.008.005.008.004.005.005.006.005.009.008.009.005.005.006.007.008.005.005.006.008.005.008.008.005.008.007.006.008.005.006.005.007.006.006.006.008.005.006.006.008.006.58

21.17

0.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.001.39

0.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.000.06

31

32

unreliable correlation between the mean values of this index with ITP the other hand WS (composed of WGS and PSS) is a too complex highlights the lack of a direct effect between the two indicators (Table and variable factor. This gives grounds to assert that the values of 5). WDLS will be influenced to a greater extent of the component whose

WDLS (Table 3) is an indicator which is formed by two variation is greater than other. Due to the lack of a direct effect on the components distinct of the effects of the pathogen – WS and LS indicator WDLS it should be taken into account the standard (Stoyanov, 2014b). For this reason, it is very difficult to be deviation of the WS or the values of WGS. This hypothesis, established a direct evaluation of the effect of black rust on it. On the combined with a unreliable correlation and the complete lack of one hand, LS is not susceptible to the effects of the disease but on significance give raise to claim that this indicator is not relevant to the

Figure 1. Spikes of Triticum polonicum accessions, damaged in different level by the pathogen of black rust: TRI933 (A); TRI3597 (B); TRI6992 (C); TRI9650 (D); TRI28851 (E); TRI28852 (F).

33

pathogen attack (Table 5). correlate significantly, though with not very high values o f the The average weight of spikelet (Table 3) is a directly correlation coefficient (Table 5). Since heredity and environmental

proportional indicator to WS, but also inversely proportional to NSS factors also influence the formation of grain and put limits on the (Stoyanov, 2014b). Again, the combination of two indicators that values of this index, the direct effect of pathogenic effect is very small have different attitudes towards ITP and the impact of the pathogen, (0.03) (Table 6). The indirect effect is significantly larger due to the are not able to give a clear vision on the changes of spikes direct effect on WGS and NGS independently from the indirect morphology. Too identical values of the standard deviation for most mechanism of action of the pathogen on the formation and of the accessions, as well as slight variations in the average values development of the grain.are not prerequisite an indirect effect on the indicator to be sought. The ratio of WGS and WS which forms grain index (Table 4) is On the other hand, the dry biomass of a single spikelets ranges an indicator of spike seedset level by weight. Therefore the impact of depending on hereditary factors, nutrients, regardless of the grains disease on fertility of the plants would have demonstrated too high. formation therein. Large and heavy glumes in some cases The presence of nearly 50% significant correlation (Table 5), determine the values of this indicator on accessions with a strong highlights the importance of the disease in the formation of spike attack and no grains to be higher than on accessions which have morphology. With similar values of correlation are indicators SEFI lower glumes and spikelets but with proper seedset. For this reason and SEFI2. Since they are corrected values of M1000 (Table 4), and because of insignificant correlation with the average values thereby to some extent neutralize the hereditary factors, the values (Table 5), the influence on spike morphology expressed by this of correlation coefficient are also over 0.4 with less than 1% indicator could not be clearly determined. significance. However, it is not possible to neutralize components

The weight of 1000 grains (Table 3) as complex indicator is too such as mineral nutrition and climate, which is the reason the direct complicated to be analyzed according to its components. However, effect of the disease on these two indicators to be unreliable. The as it gives a direct vision on the grain size (Stoyanov, 2014b), indirect effect is significantly larger (Table 6), which again highlights through it effectively could be observed a direct effect of black rust the importance of indirect effects of the pathogen on the disease on reproductive capabilities of attacked spikes. It has been reproduction apparatus of the plants, by inhibiting the active reported in other wheat species, that the grain size is greatly vegetative growth.influenced by the virulence and the aggressiveness of the pathogen All studied accessions on the basis of a three-level used scale race (Villareal et al., 1991; Beteselassie et al., 2007a; Beteselassie (Stoyanov, 2012a) are classified as susceptible or moderate et al., 2007b; McVey, 1991; Haile et al., 2012; Haile et al., 2013; susceptible to the pathogen of black rust, excluding accession Singh et al., 2006; Singh et al., 1998). Similar data were reported TRI3311, which is classified as moderate resistant. The other aside from black rust, also in brown and yellow rust (CYMMIT, 1988; accessions exhibit some distortions in the morphology of the spikes, Ivanova, 2012), powdery mildew (Bennet, 1984), septoria leaf blight particularly in the indicator M1000, which determines their (Eyal et al., 1987). All these diseases are obligate in nature, which unsuitability to participate in direct crosses to improve tetraploid determines their importance to reduce the photosynthetic activity of wheat species. Nevertheless, the creation of synthetic hexaploid leaves, stems and spikes, and hence growth inhibition in severe lines including wild species Aegilops tauschii and increasing the infestations. This leads on the one hand to a reduction of the amount resistance of the accessions to the pathogen of black rust and also of nutrients, which are intended for direct growth and reproduction of triploid wheat-bridge creation (Gerechter-Amitai et al., 1971) would plants, due to their extraction from an obligate parasitic pathogen allow the transfer of the valuable quality properties into common mycelium (Eyal et al., 1987). This leads to the formation of smaller winter wheat.grains with a low absolute weight. Therefore, average M1000

Table 5. Correlations between spikes properties and ITP of studied Triticum polonicum accessions

NSS LS LSA WS WGS NGS AI PSS NSLS WDLS WSS M1000 GI SEFI SEFI2

ITP

Pearson Correlation

.322

.020

.042

.768

-.181

.199

-.076

.591

-.209

.137

.153

.279

-.057

.690

.250

.073

.264

.058

.009

.947

-.228

.103

-.442**

.001

-.487**

.000

-.460**

.001

-.407**

.003Sig. (2-tailed)

Table 6. Total, direct and indirect effects of ITP on studied spike properties of Triticum polonicum accessions

TE – total effects, STE – standardized total effects, DE – direct effects, SDE – standardized direct effects, IE – indirect effects, SIE – standardized indirect effects

TE STE DE SDE IE SIE

NGS

WGS

PSS

NSS

M1000

SEFI

SEFI2

0.691

-0.073

0.000

0.000

-3.971

-5.077

-0.293

0.153

-0.209

0.000

0.000

-0.282

-0.219

-0.161

0.691

-0.073

0,000

0.000

-0.473

0.095

0.020

0.153

-0.209

0.000

0.000

-0.034

0.004

0.011

0.000

0.000

0.000

0.000

-3.498

-5.171

-0.313

0.000

0.000

0.000

0.000

-0.249

-0.223

-0.171

34

landrace populations from two regions of Ethiopia. Genetic ConclusionResources and Crop Evolution, 53, 387-393.Eyal Z, Scharen AL and Prescott JM, 1987. The Septoria diseases The studied accessions demonstrates a wide variation in of wheat. Concepts and methods of disease management. relation to all the parameters of the morphology of the spikes, which CYMMYT.is due to different degrees of their hereditary characteristics, mineral Ganusheva N, Yancheva S and Marcheva M, 2013. Guide in nutrition, the climatic conditions and the influence of the black rust breeding, seed production and plant biotechnologies training attack. The highest infectious type of damage exhibit accessions cources. Agricultural University Plovdiv.TRI1896, TRI3440 and TRI3442, and the lowest – accessions Gerechter-Amitai ZK, Wahl I, Vardi A and Zohary D. 1971. TRI3311. The highest values of correlation and total effect on the Transfer of stem rust seedling resistance from wild diploid einkorn to their formation by ITP are for the indicators M1000, GI, SEFI and tetraploid durum wheat by means of a triploid hybrid bridge. SEFI2, and the highest direct effect are for indicators WGS and NGS Euphytica, 20, 281-285.due to the impact of the pathogen of black rust on the formation and Haile JK, Hammer K, Badebo A, Nachit MM and Roeder MS, growth of the grains in the studied accessions. The reported total, 2012. Genetic diversity assessment of Ethiopian tatraploid wheat direct and indirect effects on the examined morphological landraces and improved durum wheat varieties using microsatellites parameters of the spikes emphasize the indirect mechanism of and markers linked with stem rust resistance. Genetic Resources action of the pathogen of black rust on reproductive organs of the and Crop Evolution, DOI 10.1007/s10722-012-9855-1.plants, by suppressing the plant growth, rather than by direct Haile JK, Hammer K, Badebo A, Singh RP and Roeder MS, 2013. damage. All studied accessions except TRI3311, are unsuitable for Haplotype analysis of molecular markers linked to stem rust participation in crosses with other tetraploid species, but are a good resistance genes in Ethiopian improved durum wheat varieties and source of genes that make them indispensable initial material to tetraploid wheat landraces. Genetic Resources and Crop Evolution, create synthetic hexaploid wheat species that could be included in 60, 853-864. the bread wheat breeding programs.Ivanova V, 2012. Studies on resistance to common wheat and other species to the cause of brown rust Puccinia triticina Erikss. Thesis for PhD, Dobrudzha Agricultural Institute, General Toshevo (Bg).AcknowledgementKang HY, Wang Y, Yuan HJ, Jiang Y and Zhou YJ, 2008. A new synthesized 6x-wheats, derived from dwarfing Polish wheat The present study was conducted with the support of IPK-(Triticum polonicum L.) and Aegilops tauschii Cosson. International Gatersleben, where from the seeds of the studied accessions were Journal of Agricultural Research, 3, 252-260.kindly provided.McVey DV, 1991. Reaction of a group of related wheat species (AABB Genome and an AABBDD) to Stem Rust. Crop Sciences, 31, 1145-1149.

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Ayala FJ and Kiger JA, 1987. Modern genetics. Zemizdat, Sofia Nikolova E and Panayotov I, 2008. Inheritance length of spike in (Bg). hybrids of winter common wheat. Proceedling of the International Bennet FGA, 1984. Resistance to powdery mildew in wheat: a Scientific Conference, 2008, Union of Scientist, Stara Zagora, 1-6 review of its use in agriculture and breeding programmes. Plant (Bg).Pathology, 33, 279-300. Pinthus MJ and Millet E, 1978. Interactions Among Number of Beteselassie N, Fininsa C and Badebo A, 2007a. Sources of Spikelets, Number of Grains and Grain Weight in the Spikes of resistance to stem rust (Puccinia graminis f. sp. tritici) in Ethiopian Wheat (Triticum aestivum L.). Annales Botanici Fennici, 42, 839-tetraploid wheat accessions. Genetic Resources and Crop 848.Evolution, 54, 337-343. Pfender WF, 2004. Role of Phenology in Host Susceptability and Beteselassie N, Fininsa C and Badebo A, 2007b. Sources of stem within-plant spread of stem rust during reproductive development of rust resistance in Ethiopian tetraploid wheat accessions. African perennial ryegrass. Phytopathology, 94, 308-316.Crop Science Journal, 15, 51-57. Rachovska G and Uhr Zl, 2010. Inheritance of quantitative CIMMYT, 1988. Breeding Strategies for Resistance to the Rusts of characteristics associated with productivity of F1 hybrids winter Wheat. Mexico. D.F. CIMMYT. common wheat, Field Crops Studies 6, 361-367 (Bg).Cox TS, Shroyer J, Ben-Hui L, Sears RG and Martin TJ, 1988. Rodriguez-Quijano M, Lucas R and Carillo JM. 2003. Waxy Genetic Improvement in Agronomic Traits of Hard Red Winter Wheat proteins and amylase content in tetraploid wheat Triticum dicoccim Cultivars 1919 to 1987. Crop Sciences, 28, 756-760. Schubl, Triticum durum L. and Triticum polonicum L. Euphytica, 134, Dochev V, Iliev I, Ivanova V and Atanasov A, 2009. Resistance to 97-101.some diseases of distributed winter common wheat varieties Roelfs AP, Singh RP and Saari EE, 1992. Rust Diseases of Wheat: (Triticum aestivum L.). Filed Crop Studies, 5, 39-44. Concepts and methods of disease management. Mexico, D.F.: Dorofeev VF, 1987. Pšenica polonicum (T. polonicum L.). In: CIMMYT.Dorofeev VF (ed) Pšenicy mira: Vidovoj sostav, dostiženiâ selekcii, Roelfs AP, 1988. Genetic control of phenotypes in wheat stem rust. sovremennye problemy i ishodnyj material, 2nd edn. Leningrad, Annual Review of Phytopathology, 26, 351-367.Kolos, 48-50. Singh RP, Hodson DP, Jin Y, Huerta-Espino J, Kinyua MG, Elbaum R, Zaltzman L, Burgert I and Fratzl F, 2007. The Role of Wanyera R, Njau P and Ward RW, 2006. Current status, likely Wheat Awns in the Seed Dispersal Unit. Science, 316, 884-886. migration and strategies to mitigate the threat to wheat production Eticha F, Belay G and Bekele E, 2006. Species diversity in wheat from race Ug99 (TTKS) of stem rust pathogen. CAB Reviews:

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Review

Genetics and Breeding

Nutrition and Physiology

Production Systems

Molecular mechanisms and new strategies to fight stresses in egg-producing birds E. Shatskikh, E. Latypova, V. Fisinin, S. Denev, P. Surai

Gene action in the inheritance of date to ear emergence and time to physiological maturity in bread wheat crosses (Triticum aestivum L.)N. Tsenov, T. Gubatov, E. Tsenova

Productivity and stability of the yield from common winter wheat cultivars developed at IPGR Sadovo under the conditions of Dobrudzha region P. Chamurliyski, E. Penchev, N. Tsenov

Effectof black (stem) rust (Puccinia Graminis F.SP. Tritici) attack to the spike characteristics in Polishwheat (Triticum Polonicum L.)H. Stoyanov

Analysis of DNA polymorphism of CAST gene in Local Karnobat and Stara Zagora sheep breedsD. Hristova, S. Georgieva, S. Tanchev

Correlation between grain yield and yield components in winter barley varieties N. Markova Ruzdik, D.Valcheva, D.Vulchev, Lj. Mihajlov, I. Karov, V. Ilieva

Genetic diversity in different accessions of oat (Avena sativa L.)T. Savova, B. Dyulgerova, G. Panayotova

Interspecific hybridization in cotton and its use in breedingA. Stoilova, I. Saldzhiev

Influence of the direction of crossing on heterosis and transgression events in relation to the length of the vegetative period of Burley tobaccos variety group Y. Dyulgerski, T. Radoukova, L. Dospatliev

The performance of female dairy calves fed texturized starters with different protein sources E. Yavuz, N. Todorov, G. Ganchev, K. Nedelkov

Feeding value estimation of spring forage pea (Pisum sativum L.) in organic cultivationI. Nikolova, N. Georgieva, Y. Naydenova

Treatment of post harvest residues with cellulose decomposing preparationsI. Effect on grain yield from wheatG. Milev, I. Iliev, A. Ivanova

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Seasonal dynamics of important for Coriandrum sativum virus pathogensB. Dikova, H. Lambev

Crop relationship "yield-evapotranspiration" for green beanR. Kalaydzhieva, D. Davidov, A. Matev, V. Kuneva

Species composition and density of weeds in a wheat crop depending on the soil tillage system in crop rotationP. Yankov, M. Nankova, M. Drumeva, D. Plamenov, B. Klochkov

Assessment of Bulgarian Black Sea coastal water using the biological quality element phytoplanktonD. Petrova, D. Gerdzhikov

Evaluation on reaction of late maturing maize hybrids and lines to Fusarium ear rotM. Haddadi, M. Zamani

Contemporary state of macrophytobenthos along the Bulgarian coast of the Black SeaD. Petrova, V. Vachkova, D. Gerdzhikov

Bioconversion of nitrogen in eco-technical system for eggs productionA. Gencheva

Mixed viral infections in tomato as a precondition for economic lossN. Petrov

Storage and its effect on the antioxidant capacity of dried Bulgarian Chrysanthemum balsamita L.A. Popova, D. Mihaylova, I. Alexieva

Correcting the breadmaking quality of flour damaged by Sunn pest (Eurygaster integriceps) by using apple pectinI. Stoeva

Investigation the influence of dietary fiber on the rheological properties of alginate beadsZ. Manev, N. Petkova, P. Denev, D. Ludneva, S. Zhelyazkov

Occurrence of Pseudomonas syringae pv. tomato in BulgariaM. Stoyanova, K. Aleksandrova, D. Ganeva, N. Bogatzevska

Agriculture and Environment

Product Quality and Safety

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Instruction for authors

Preparation of papersPapers shall be submitted at the editorial office typed on standard typing pages (A4, 30 lines per page, 62 characters per line). The editors recommend up to 15 pages for full research paper ( including abstract references, tables, figures and other appendices)The manuscript should be structured as follows: Title, Names of authors and affiliation address, Abstract, List of keywords, Introduction, Material and methods,Results, Discussion, Conclusion, Acknowledgements (if any), References, Tables, Figures.The title needs to be as concise and informative about the nature of research. It should be written with small letter /bold, 14/ without any abbreviations. Names and affiliation of authorsThe names of the authors should be presented from the initials of first names followed by the family names. The complete address and name of the institution should be stated next. The affiliation of authors are designated by different signs. For the author who is going to be corresponding by the editorial board and readers, an E-mail address and telephone number should be presented as footnote on the first page. Corresponding author is indicated with *.Abstract should be not more than 350 words. It should be clearly stated what new findings have been made in the course of research. Abbreviations and references to authors are inadmissible in the summary. It should be understandable without having read the paper and should be in one paragraph. Keywords: Up to maximum of 5 keywords should be selected not repeating the title but giving the essence of study. The introduction must answer the following questions: What is known and what is new on the studied issue? What necessitated the research problem, described in the paper? What is your hypothesis and goal ?Material and methods: The objects of research, organization of experiments, chemical analyses, statistical and other methods and conditions applied for the experiments should be described in detail. A criterion of sufficient information is to be possible for others to repeat the experi-ment in order to verify results.Results are presented in understandable

tables and figures, accompanied by the statistical parameters needed for the evaluation. Data from tables and figures should not be repeated in the text.Tables should be as simple and as few as possible. Each table should have its own explanatory title and to be typed on a separate page. They should be outside the main body of the text and an indication should be given where it should be inserted.Figures should be sharp with good contrast and rendition. Graphic materials should be preferred. Photographs to be appropriate for printing. Illustrations are supplied in colour as an exception after special agreement with the editorial board and possible payment of extra costs. The figures are to be each in a single file and their location should be given within the text. Discussion: The objective of this section is to indicate the scientific significance of the study. By comparing the results and conclusions of other scientists the contribution of the study for expanding or modifying existing knowledge is pointed out clearly and convincingly to the reader.Conclusion: The most important conse- quences for the science and practice resulting from the conducted research should be summarized in a few sentences. The conclusions shouldn't be numbered and no new paragraphs be used. Contributions are the core of conclusions. References:In the text, references should be cited as follows: single author: Sandberg (2002); two authors: Andersson and Georges (2004); more than two authors: Andersson et al.(2003). When several references are cited simultaneously, they should be ranked by chronological order e.g.: (Sandberg, 2002; Andersson et al., 2003; Andersson and Georges, 2004).References are arranged alphabetically by the name of the first author. If an author is cited more than once, first his individual publications are given ranked by year, then come publications with one co-author, two co-authors, etc. The names of authors, article and journal titles in the Cyrillic or alphabet different from Latin, should be transliterated into Latin and article titles should be translated into English. The original language of articles and books translated into English is indicated in parenthesis after the bibliographic reference (Bulgarian = Bg, Russian = Ru, Serbian = Sr, if in the Cyrillic, Mongolian =

Мо, Greek = Gr, Georgian = Geor., Japanese = Jа, Chinese = Ch, Arabic = Аr, etc.)The following order in the reference list is recommended:Journal articles: Author(s) surname and initials, year. Title. Full title of the journal, volume, pages. Example:Simm G, Lewis RM, Grundy B and Dingwall WS, 2002. Responses to selection for lean growth in sheep. Animal Science, 74, 39-50Books: Author(s) surname and initials, year. Title. Edition, name of publisher, place of publication. Example: Oldenbroek JK, 1999. Genebanks and the conservation of farm animal genetic resources, Second edition. DLO Institute for Animal Science and Heal th, Netherlands.Book chapter or conference proceedings: Author(s) surname and initials, year. Title. In: Title of the book or of the proceedings followed by the editor(s), volume, pages. Name of publisher, place of publication. Example: Mauff G, Pulverer G, Operkuch W, Hummel K and Hidden C, 1995. C3-variants and diverse phenotypes of unconverted and converted C3. In: Provides of the Biological Fluids (ed. H. Peters), vol. 22, 143-165, Pergamon Press. Oxford, UK.Todorov N and Mitev J, 1995. Effect of level of feeding during dry period, and body condition score on reproductive perfor-

thmance in dairy cows,IX International Conference on Production Diseases in Farm Animals, September 11–14, Berlin, Germany.Thesis:Hristova D, 2013. Investigation on genetic diversity in local sheep breeds using DNA markers. Thesis for PhD, Trakia University, Stara Zagora, Bulgaria, (Bg).

The Editorial Board of the Journal is not responsible for incorrect quotes of reference sources and the relevant violations of copyrights.

Animal welfareStudies performed on experimental animals should be carried out according to internationally recognized guidelines for animal welfare. That should be clearly described in the respective section “Material and methods”.

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