Mining and validation of novel genotyping-bysequencing(GBS)-based simple sequence repeats(SSRs)and their application for the estimation of the genetic diversity and population structure of coconuts(Cocos nucifera L.)in Thailand

Coconut(Cocos nucifera L.)is an important economic crop in tropical countries.However,the lack of a complete reference genome and the limitations of usable DNA markers hinder genomic studies and the molecular breeding of coconut.Here,we present the results of simple sequence repeat(SSR)mining from a high-throughput genotyping-bysequencing(GBS)study of a collection of 38 coconut accessions.A total of 22,748 SSRs with di-,tri-,tetra-,penta-and hexanucleotide repeats of five or more were identified,2451 of which were defined as polymorphic loci based on locus clustering in 38 coconut accessions,and 315 loci were suitable for the development of SSR markers.One hundred loci were selected,and primer pairs for each SSR locus were designed and validated in 40 coconut accessions.The analysis of 74 polymorphic markers identified between 2 and 9 alleles per locus,with an average of 3.01 alleles.The assessment of the genetic diversity and genetic relationships among the 40 coconut varieties based on the analysis of population structure,principal coordinate analysis(PCoA),and phylogenetic tree analysis using the 74 polymorphic SSR markers revealed three main groups of coconuts in Thailand.The identified SSR loci and SSR markers developed in this study will be useful for the study of coconut diversity and molecular breeding.The SSR mining approach used in this study could be applied to other plant species with a complex genome regardless of the availability of reference genome.

Characterization of Myanmar Paw San Hmwe Accessions Using Functional Genetic Markers

Paw San Hmwe （PSM） rice has been cultivated in many areas of Myanmar for a long time. Strong aroma, good taste and its elongation during cooking are the key characteristics of PSM rice. Thirty-one PSM accessions were genotypically characterized, and their physical grain and cooking quality traits were studied. We used specific gene markers associated with aroma, apparent amylose content （AAC） and alkali spreading value to determine the alleles carried by different PSM accessions. The results revealed that six PSM accessions （PSM10, PSM12, PSM13, PSM21, PSM22 and PSM30） had a 3-bp insertion in Os2AP gene. Gel consistency （GC） allele was predominant among the PSM accessions for gelatinization temperature （GT）, however, the phenotype observed was between low and intermediate GT because of the combination of the GC allele with the presence of low GT allele at heterozygous state from the other loci of the SSIla gene. Intermediate to high AAC was observed among the PSM accessions corresponding to the haplotype identified for the single nucleotide polymorphism G/T and the （Cm）n repeat in the Wx gene. The characterization and grouping data of PSM accessions posted benefits to Myanmar seed banks, and our results will help in maintaining the integrity of PSM rice variety.

Evaluation of Agronomic Traits in Chromosome Segment Substitution Lines of KDML105 Containing Drought Tolerance QTL under Drought Stress

Drought is a major abiotic constraint to rice production in rainfed lowland and insufficiently irrigated areas. The improvement of drought tolerant varieties is one of the strategies to reduce the negative effects of drought. Quantitative trait loci （QTLs） for primary and secondary traits related to drought tolerance （DT） on chromosomes 1, 3, 4, 8 and 9 that determined from double haploid lines derived from a cross between CT9993 and IR62266 were introgressed and dissected into small pieces in the genetic background of Khao Dawk Mall 105 （KDML105） to develop chromosome segment substitution line （CSSL） population. The CSSLs were evaluated at the reproductive stage for their agronomic performance and yield components under drought stress, and results were compared with irrigated condition. The flowering of CSSL lines was 6 to 7 d earlier than KDML105. The mean values of grain yields in the CSSLs were higher than KDML105 under drought and irrigated conditions. At irrigated condition, the grain yields of introgression lines carrying DT-QTLs from chromosomes 4 and 8 were higher than that of KDML105, whereas other traits showed little difference with KDML105. Analysis indicated that grain yield has positive correlation with plant height, tiller and panicle number per plant, and total grain weight per plant under drought stress while negatively correlated with days to flowering. As mentioned above, CSSLs showing good adaptation under drought stress can be used as genetic materials to improve drought tolerance in Thai rainfed lowland rice breeding program, and as materials to dissect genes underlying drought tolerance.