Naghavi MR

Aghaei MJ

Taleei AR

Omidi M

Hassani ME

English

Sydney eScholarship

 Genetic diversity of hexaploid wheat and three Aegilops species using microsatellite markers  Naghavi MR 1, Aghaei MJ2, Taleei AR1, Omidi M1 and Hassani ME3 1Agronomy and Plant Breeding Dept., Agricultural College, University of Tehran, Karaj, Iran 2 Seed and Plant Improvement Institute, National Plant Gene Bank of Iran, Mahdasht Road, Karaj, Iran 3 Horticultural Dept., Agricultural College, University of Tehran, Karaj, Iran   ABSTRACT  In this study a set of 21 simple microsatellite primers were used to determine the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (DtDt), Ae. cylindrica (CCDcDc) and Ae. crassa (MMDcr1Dcr1), collected from 13 different sites in Iran.  A total of 273 alleles were detected across all four species and the number of alleles per microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Cluster analysis classified the four species in three groups, locating Ae. tauschii and Ae. cylindrica in the same cluster using the UPGMA method. It appeared that different genotypes of Ae. tauschii could be involved in the evolution of polyploid species. A high level of variation and also the highest number of unique alleles were observed within Ae. crassa accessions, indicating this species as a good potential source of novel genes for bread wheat improvement.   INTRODUCTION  Synthetic hexaploid wheat lines, produced by hybridization between commercial cultivars of T. durum or T. aestivum with Ae. tauschii, were used to improve the genetic base of hexaploid wheat (Kazi et.al., 1996). Several of these lines and synthetic derivative cultivars are known to have important agronomical traits such as pest and disease resistance, environmental stress tolerance and adaptability to varied climatic conditions. The D genome is not limited to T. aestivum and Ae. tauschii. Two tetraploid wild species, Ae. cylindrica (2n=4x=28, CCDD) and Ae. crassa (2n=4x=28, MMDD), which are widely distributed in Iran, contain the D genome as well. However, there is no evidence that these species contributed to the D genome of hexaploid wheat. The distribution of Ae. tauschii is limited to Northern Iran, while the distribution of the other two tetraploid species is wider in Northern and Western Iran.   Ae. cylindrica growns in the most suitable areas north of Iran from Khorasan to Azarbaijan provinces, west of Iran from Azarbaijan to Lorestan provinces and in the center of Iran in such places as Semnan, Tehran, Ghazvin, Ghom, Markazi, Isfahan and Fars provinces. However, Ae. crassa is not widely distributed like Ae. cylindrica, but it is distributed in west of Iran, from Azarbaijan to Bushehr provinces in south, and central provinces such as Tehran, Ghazvin, Ghom, Markazi, Isfahan and Fars. Ae. crassa does not occur in Northern Iran, which is the main distribution area of Ae. tauschii. Introduction of different adaptability genes from these tetraploid species to bread wheat will produce new cultivars, with wider genetic base and adaptability, if the diversity of the D genome in these species is different to that in Ae. tauchii and T. aestivum.   Wide diversity within the D genome of Ae. tauschii populations in contrast with the Dt genome of T. aestivum was confirmed by an investigation with AFLP molecular markers (Dvorak et al., 1998, Lelly et al., 2000, Pestosova et al., 2000).   Genetic diversity in this species was evaluated using allozyme diversity (Watanabe et al. 1994), RAPD molecular markers (Goryunova et al. 2004), a combination of RAPD and AFLP molecular markers (Pester et al. 2003) and polymorphism for DNA sequences (Caldwell et al. 2004). Genetic diversity in the tetraploid species, Ae. crassa, was evaluated using isozyme markers (Nakai, 1989), SSR markers (Zhang and Devorak 1992; Naghavi et al. 2007) and in situ hybridization (Badaeva et al. 1998). The potential of introducing agronomic traits such as cytoplasmic male sterility (Murai  et al. 1993), yield, seed quality and salinity tolerance (Liu et al. 2002) from Ae. crassa into wheat breeding programs has been demonstrated by various researchers.  Populations of four species with the D genome from the  National Plant Gene Bank of Iran were studied using microsatellite markers to evaluate genetic diversity among and within populations and to determine relationships between the D genome of these species.  MATERIALS AND METHODS  Fifty two accessions from four species, T. aestivum, Ae. tauschii, Ae. cylindrica and Ae. crassa, provided from the National Plant Gene Bank of Iran were used in this study. The accessions were collected from 13 different sites (for each species and from each site one accession was selected) in Iran. The sites were considered as centers of diversity for those species in Iran and usually most of the species are found to be prevalent there. The accessions were planted under  greenhouse conditions and genomic DNA was extracted from young leaves according to the method of Saghaei-Maroof et al. (1984). A total of 21 primer pairs (Roder et al. 1998), at least one primer pair from each arm of seven chromosomes of D genome, were selected for genotyping assays. The total number of alleles and polymorphic alleles were counted using PopGene 3.2. Polymorphism Information Content (PIC) was determined for each primer pair.  RESULTS AND DISCUSSION  A total of 272 alleles were detected by 21 primers. The number of alleles per primer ranged from 3 for WMS111 to 27 for WMS314 with an average number of 13 alleles per locus (Table 1). The lowest and highest PIC was 0.283 and 0.728 to WMS232 and WMS 311 respectively. Lelley et al. (2000) observed 90 different alleles for 14 microsatellite primers in Ae. tauschii populations. Their average of PIC was 0.68, which is a little larger than ours. Pestova et al. (2000) found 11-25 and an average of 18.8 alleles per microsatellite in evaluation of Ae. tauschii populations.  They observed 338 alleles for 18 primers, and the heterogeneity index varied from 0.75 to 0.95. In the evaluation of the genetic diversity of Iranian Ae. tauschii populations using 9 microsatellites, a total of 66 alleles and 4-12 alleles per primer was observed. The average of PIC was 0.659 (Saeidi et al. 2005). These results were similar to that of other researchers. Any disparities were mainly related to the differences in primers used and the very diverse material (genotypes) used. A total of 272 alleles were detected by 21 primers. The number of alleles per primer ranged from 3 for WMS111 to 27 for WMS314 with an average number of 13 alleles per locus.  The average of Nei’s index of genetic diversity was 0.5008 among the four species. The highest diversity was 0.5571 in Ae. tauschii and the lowest diversity, 0.4697, was found in Ae. cylindrica (Table 2). In polyploid species that derived their D genome from Ae. tauschii the highest diversity was found in Ae. carassa and lowest diversity in Ae. cylindrica. The genetic diversity of T. aestivum was between that of Ae. carassa and Ae. cylindrical, but was more similar to Ae. cylindrica.   Genetic variability reported in Dt genome of bread wheat is much smaller than Ae. tauschii according to seed storage proteins, allozymes and DNA loci (Murphy et al. 1997). Low diversity in Ae. cylindrica populations were found based on  variability of allozymes (Watanabe et al. 1994), C-banding (Badaeva et al. 2002), RAPD markers (Goryunova et al. 2004), a combination of RAPD and AFLP markers (Pester et al. 2003) and polymorphism for DNA sequences (Caldwell et al. 2004).   It is suggested that Ae. cylindrica is a relatively new tetraploid species (Gandhi et al. 2005). Some results suggested that Dt genome of bread wheat and Dc genome of Ae. cylindrica inherited from different biotype of Ae. tauschii (Badaeva et al. 2002; Caldwell et al. 2004). However, the relationship between Ae. cylindrica and subspecies of Ae. tauschii is not clearly understood (Gandhi et al. 2005). C-banding and in-situ hybridization studies on a tetraploid biotype of Ae. crassa suggested that the Dcr1 genome of 4x Ae. crassa is highly modified compared with the D genome of the progenitor species Ae. tauschii (Badaeva  et al. 1998).   Table1: List of 21 wheat microsatellites, the number of alleles and PIC Microsatellites Number of alleles PIC WMS484 19 0.664WMS157 6 0.524WMS469 14 0.633WMS383 19 0.662WMS337 14 0.644WMS642 13 0.597WMS190 23 0.574WMS192 14 0.575WMS194 18 0.605WMS261 16 0.660WMS314 27 0.715WMS161 9 0.679WMS295 10 0.688WMS111 3 0.353WMS212 11 0.649WMS37 9 0.677WMS311 21 0.728WMS624 12 0.651WMS232 4 0.283WMS114 6 0.473WMS33 5 0.298  Table 2- The Nei’s index of genetic diversity for species Species No. Genetic diversity Index T. aestivum 13 0.4797 Ae. crassa 13 0.4966 Ae. cylindrica 13 0.4697 Ae. tauschii 13 0.5571  In conclusion, this study confirms the usefulness of SSR markers to study wheat genetic diversity. Additionally, the results obtained from this study could be useful for improving the understanding of diversity in and management of germplasm collections.  REFERENCES  1. Badaeva E.D., Friebe B., Zoshchuk S.A., Zelenin A.V., Gill B.S. 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Genetic diversity of hexaploid wheat and three Aegilops species using microsatellite markers

Oort, J. van

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Het Evangelie van Judas en de Joods-christelijke dialoog

https://ses.library.usyd.edu.au/bitstream/2123/3231/1/P028.pdf

Genetic diversity of hexaploid wheat and three Aegilops species using microsatellite markers

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