Geographical variation of foxtail millet, Setaria italica (L.) P. Beauv. based on rDNA PCR–RFLP

The rDNA PCR–RFLP of foxtail millet (Setaria italica) germ-plasm collected throughout Eurasia and from a part of Africa was investigated with five restriction enzymes according to our previous study. Foxtail millet germ-plasms were classified by length of the rDNA IGS and RFLP; clear geographical differentiation was observed between East Asia, the Nansei Islands of Japan-Taiwan-the Philippines area, South Asia and Afghanistan-Pakistan. We also found evidence of migration of foxtail millet landraces between the areas. We calculated diversity index (D) for each region and found that center of diversity of this millet is East Asia such as China, Korea and Japan.     

rDNA polymorphism of foxtail millet (Setaria italica ssp. italica) landraces in northern Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis)

Ribosomal DNA (rDNA) spacer length polymorphism was studied in foxtail millet (Setaria italica ssp. italica) landraces from Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis) from Pakistan by PCR-based methods. Sequence polymorphism was also investigated for accessions selected based on the observed length polymorphism. The PCR-based length polymorphism and sequence polymorphism of rDNA intergenic spacer (IGS) clearly demonstrated genetic differentiation between cultivated and wild forms in the region. Genetic differentiation was observed between different areas to some extent in the cultivated form, and between different regions in the wild form of northern Pakistan. Based on the results, we discuss the genetic differentiation of foxtail millet and wild ancestor in this region and possible utility of rDNA markers to trace the dispersal of this crop in the region.     

Origin of waxy common millet, Panicum miliaceum L. in Japan

We developed PCR-based markers for genotyping Waxy genes in common millet (Panicum miliaceum L.) and proposed a new hypothesis on the origin and dispersal of waxy type of Japanese landraces. I₂-K staining indicated that almost all the Japanese landraces had waxy endosperm. Based on the PCR and dCAPs analyses of two waxy loci on different genomes we concluded that there were at least two possible lineages in Asian continent that had independent dispersal pathways into Japan. Most Japanese common millet had the similar genotypes with those in Korean Peninsula but a few particularly from northern part of Japan had common genotypes in northeastern China and Primorskaya Province of Russia.     

A phenotypic diversity analysis of foxtail millet (Setaria italica (L.) P. Beauv.) landraces of Chinese origin

A total of 23 381 foxtail millet landraces of Chinese origin were analysed for seven qualitative traits and four quantitative traits. The Shannon-Weaver diversity index was used to estimate the phenotypic diversity of each characteristic on the basis of administrative provinces and ecogeographical regions. Hierarchical analysis of variance indicated that most of the variation was due to differences among characteristics. Only the diversity indices for leaf color of seedlings, starch composition and 1000-grain weight showed significant differences among regions. In relative terms, a greater genetic divergence was found in some provinces of southern China, where foxtail millet is not a major crop.     

Geographical variation of nuclear genome RFLPs and genetic differentiation in foxtail millet, Setaria italica (L.) P. Beauv.

Sixteen RFLP loci in 62 landraces were assayed to study geneticdifferentiation in foxtail millet, Setariaitalica (L.) P. Beauv. Among 52 bands, 47 werepolymorphic among foxtail millet landraces. A dendrogram constructedbased on RFLPs was divided into five major clusters (clusterI–V). Clusters I and II contained strains mainly from EastAsia. Cluster III consisted of strains from subtropical and tropicalregions in Asia such as Nansei Islands of Japan, Taiwan and thePhilippines and India and cluster IV consisted of some strains fromEast Asia, a strain from Nepal and a strain from Myanmar. Cluster Vcontained strains from central and western regions of Eurasia such asAfghanstan, Central Asia and Europe. Chinese landraces wereclassified into four clusters. These results indicate that foxtailmillet landraces have differentiated genetically between differentregions and that Chinese landraces were highly variable.     

Genetic diversity analysis of yams (Dioscorea spp.) cultivated in China using ISSR and SRAP markers

Yam (Dioscorea spp.) is widely cultivated in China and many landraces are maintained by local farmers. However, there is little information available about their diversity and species identity. In this study, inter simple sequence repeat (ISSR) and sequence related amplified polymorphism (SRAP) techniques were used to assess genetic diversity within 21 yam landraces from seven cultivated populations. We observed high level of polymorphism among these landraces, specifically, 95.3 % for ISSR and 93.5 % for SRAP. Analysis of molecular variance revealed a significantly greater variation among the four yam species (40.39 %) and their populations (35.78 %) than within the populations (23.83 %). The unweighted pair group method arithmetic averages clusters and principal component analysis for 21 landraces formed four well-separated groups containing landraces of each of the four species, namely, Dioscorea opposita Thunb., Dioscorea alata L., Dioscorea persimilis Prain et Burkill, and Dioscorea fordii Prain et Burkill. The ISSR and SRAP primers were highly discriminatory among the 21 landraces; all 21 landraces could be easily differentiated using these primers. The average mean of gene flow (Nm = 0.1081) estimated from high genetic differentiation (Gst = 0.8222) suggested that gene flow among the populations was relatively restricted. The lack of genetic diversity within individual yam species suggests that it is critical to develop long-term strategies for enhancing genetic diversity within various yam species.     

Molecular diversity of restriction enzyme sites, Indels and upstream open reading frames (uORFs) of 5′ untransalted regions (UTRs) of Waxy genes in Triticum L. and Aegilops L. species

5′ Untransalted regions (UTR) sequences of Waxy genes were amplified from all 81 Triticum L. and Aegilops L. species by PCR with specific primers. It was found that the sequence length at 7D loci was longer than that 7B and 7A. These sequences contained 170 singleton variable sites and 484 polymorphic sites and that the average length of Indels was 8.5?bp. There were abundant regions of restriction enzyme sites and two regions of simple sequence repeat, “GAA” and “CTGA”, in all sequences. A total of 65 uORFs were detected and classed into 37 types, with the variation in uORFs mainly due to single nucleotide polymorphisms (SNPs), and also to the presence of Indels. All sequences in tetraploids and hexaploids could be grouped into Types A, B, I, II, GI and GII based on sequence variation. Restriction enzyme sites, Indel polymorphisms and the classes of uORFs present together indicated that Type I was more similar to Ae. tauschii whereas Type II was more similar to Type B, and both more similar to Ae. longissima; Type A was more similar to Ae. speltoides. Population analysis was performed and Neighbour-joining trees derived from different species, types and accessions further confirmed that the ancestors of T. urartu, Ae. speltoides, Ae. longissima and Ae. tauschii were involved to the evolution of common wheat, and also implied that Ae. longissima might have participated later than Ae. speltoides. 5′ UTRs of Waxy genes in tetraploid and hexaploid species conserved characters from their respective progenitors when compared with diploid. For the first time, we are able to conclude there is abundant variation in SNPs, Indels and uORFs between 5′ UTRs of Waxy genes from different species related to common wheat, and suggest that further research could help to understand Waxy gene function more deeply and hence improve wheat breeding. Our results also show that three hexaploid species in China have unique diversity in the 5′ UTR of their Waxy gene.     

Physical and Molecular Characterization of Millet Starches

This study investigated the physical and molecular starch characteristics of four Canadian‐grown millet species: pearl, foxtail, proso, and finger millet. The millet starch granules ranged from about 2.5 to 24 μm in size and were mainly polygonal with a few spherical ones. Their amylose contents ranged from 28.6 to 33.9%, with finger and pearl millets having much more of long amylose chains than short amylose chains compared with foxtail and proso millets. Starches also differed in the molecular structure of their branched amylose, with finger and pearl millets having longer glucan chains between branch points. The enthalpy of gelatinization of starch granules ranged from 11.8 to 13.2 J/g, and the enthalpy of melting of the retrograded starches ranged from 2.2 to 5.9 J/g. The onset temperature of gelatinization (T_o) of the starches ranged from 62.8 to 70.6°C. Addition of iodine vapor to the granular starches showed significant (P < 0.05) differences in the ratio of the absorbance to scattering coefficient (K/S) values, indicating differences in the rigidity of the glucan chains present in the granules. Starches with short amylose chains exhibited higher K/S values. Iodine vapor addition resulted in altered X‐ray diffractogram peak intensities. The study suggested differences in the structure and granular architecture of the millet starches.     

Genetic diversity among Polish landraces of common oat (Avena sativa L.)

All over the world about 220,000 accessions of the genus Avena is preserved in gene banks. Polish oats collection, in the National Centre for Plant Genetic Resources, consists of about 2,500 accessions. More than 80 % of them belong to the species Avena sativa. Only 136 accessions have the landrace or traditional cultivar status and 91 of them were collected in Poland. The main objective of this study was to estimate genetic diversity of a set of 67 Polish landraces using inter simple sequence repeat (ISSR) markers. We also tried to determine whether genetic diversity depended on the region and altitude of collection site and if there was a relationship between DNA polymorphism and some morphological traits. The accessions were collected between years 1973 and 1999 from areas of Northern, Southern and Eastern Poland. The obtained results indicated a relatively low diversity (0.09–0.37) of the common oat landraces. Analysis of molecular variation (AMOVA) showed that there were no significant differences within designated geographic regions. It indicated that at the same time the altitude of collection site had significant effect on the genetic differentiation of the accessions. AMOVA also showed the presence of variance between groups formed on the basis of the colour of lemma. White lemma accessions were also characterized by highest genetic distinctiveness. Considering the evident impact of local climatic conditions on the genetic distinctiveness of Polish landraces utility of them in the Polish breeding programs should be considered.     

Development of EST-SSR in foxtail millet (<Emphasis Type="Italic">Setaria italica</Emphasis>)

The development of EST-SSR in foxtail millet (Setaria italica) for polymorphism and transferability study was reported here. From 1213 EST sequences, 30 SSRs were obtained and primers were designed for 26 SSRs. Among them, four pairs of SSR primers amplified polymorphic products in 12 foxtail millet cultivars and one accession of Setaria viridis, a wild relative of foxtail millet, with 10 alleles detected for the four loci and 2.5 alleles per locus. In addition, ten SSR markers could be transferred to other nine Gramineae species. The putative functions of 11 ESTs containing polymorphic and transferable SSRs were also identified.     

Ecological stability of genetic diversity among landraces of little millet (<Emphasis Type="Italic">Panicum sumatrense</Emphasis>) in south India

Risk-prone agriculture by resource-poor tribal farmers in Kolli hills, south India is slowly giving way to commercial exploitation for paltry compensation thus eroding genetic diversity of millets. Cultivation and conservation of millets, particularly, little millet (Panicum sumatrense Roth ex Roem. et Schult.), called samai in the local language, Tamil are on the wane. Whether irregular and sparse cultivation of little millet by tribals has an impact on the current level of genetic divergence is also not clear. Therefore genetic divergence among 7 landraces and 1 check variety was evaluated based on morphometric traits at two sites over two seasons. The seven landraces formed six distinct groups with high inter-group distances. Days to maturity and flowering time contributed the most to genetic differentiation. Principal component analysis confirmed the results of divergence analysis. But a recent molecular analysis of diversity reported that the landraces were all genetically uniform and any observed diversity could be due environmental variation. An analysis of the contradictory results only emphasized the fact that lack of polymorphism need not imply lack of genetic divergence. The findings co evaluated with relevant published work highlight the value of morphometric analysis and reveal sustained genetic divergence in little millet.     

Extensive introgression of Middle American germplasm into Chilean common bean cultivars

Summary The genetic diversity of 95. representative Chilean common bean (Phaseolus vulgaris L.) landraces was analyzed using phaseolin seed protein and eight isozyme systems as genetic markers. Four types of phaseolin were found, “C”, “T”, “S” and “H”, in decreasing order of frequency. Each type had a different distribution between the Northern and Southern regions of the country. Nei’s genetic distance based on isozyme diversity indicated that a high percentage of the total variation found in this sample occurred between landraces and only a small percentage of the variation was detected within populations. Cluster analysis based on Nei’s genetic distance and a principal component analysis of isozyme frequencies did not detect a clear association between the geographic distribution of the landraces and their isozyme constitution. However, Nei’s genetic distance analysis clustered the bean landraces into two major groups which had a specific isozyme pattern, seed color, and seed size. The genetic analysis also detected a rare polymorphism for theMdh-2 locus, a null allele at theDiap-2 locus, and polymorphism for theAco-2 locus. The principal component analysis of isozyme frequencies showed that only 30% of the genotypes analyzed were similar to the Andean check and 5% of the samples were similar to Middle American check. This finding suggests a high frequency of hybridization between the Middle America and Andean gene pools in cultivated common bean from Chile.     

Molecular and Chemical Characterization of a New Waxy Allele in Barley (Hordeum vulgare L.)

Barley m38 mutant was selected for its high level of mixed‐linkage (1,3),(1,4)‐β‐d ‐glucan (MLG) in the grain. This elevated level of MLG was found to be associated with decreased amylose accumulation as well as other chemical composition alterations. Molecular characterization results revealed m38 as a new allele of the Waxy gene, encoding an endosperm‐specific granule‐bound starch synthase I (GBSSI). Additional mapping data from amylose phenotype and GBSSI gene specific markers supported the conclusion of the GBSSI mutation in m38. The m38 locus contains a nucleotide alteration that would result in the substitution of glycine at position 263 with serine in the putative adenosine‐5′‐diphosphate‐glucose binding domain. This amino acid substitution alters loop structures on the exterior surface of the folded protein and may affect its enzyme activity. Characterizations of m38 in this report provide for a new allele of the Waxy gene and additional evidence of pleiotropic effects on other chemical components including increased MLG, fructans, and fats and decreased amylose and protein.     

不同生态区骨干谷子品种表型鉴定与遗传分析

骨干谷子品种对谷子产业发展发挥了重要的作用,为了解其优异性状遗传基础及利用价值,选用来源于不同生态区的12个骨干谷子品种,进行表型鉴定和遗传差异分析。结果表明,12个来源于不同生态区的骨干品种在株高、穗长、穗粗、穗下节间长、单穗重、生育期以及黄色素含量等方面差异较大。79对SSR多态性标记共检测到258个多态性变异位点,平均3.265 8个;SSR标记多态性(PIC)的变幅为0.141 1~0.711 6,平均为0.510 1;其中PIC>0.5的SSR标记有50对,占多态性标记的63.3%。12份材料的遗传距离变幅为0.140 2~0.801 1,平均为0.473 7;来自华北夏谷区的豫谷1号和沧谷4号遗传距离最近,而沧谷4号和来自内蒙高原的地方品种金香玉距离最远;总体而言,华北夏谷区品种和西北早熟春谷区品种遗传距离相对较大。相似性系数为0.594时,12个品种可聚为两类,华北夏谷区的豫谷1号、矮88、冀谷19号、沧谷4号和济谷12聚为一类,来自西北春谷早熟区、晚熟区和东北春谷区的7个品种聚为一类,聚类分析结果和品种的生态类型具有较高的一致性。群体结构分析和聚类分析的结果相似,华北夏谷区品种和春谷区品种之间存在基因交流。总之,不同生态区来源品种间遗传差异较大,同一生态区来源品种遗传差异较小;华北夏谷区骨干品种之间遗传差异较春谷区品种遗传差异相对较小;华北夏谷区品种和西北早熟春谷区品种间遗传差异较大,丰富华北夏谷区遗传变异,应加强与西北早熟春谷区品种之间的交流。本研究结果为优异谷子品种培育及种质资源利用提供了一定依据。     

谷子EMS矮秆突变体的产量性状分析

为丰富谷子遗传多样性和突变体库材料,本研究采用1%浓度的甲基硫磺乙酯(EMS)诱变晋谷40号 10 h, 在谷子生育期间对突变体M1和M2代材料进行表型性状调查。结果表明,共获得332株表型差异突变型单株,这些突变体类型主要包括株高、穗型、茎粗、育性、刚毛、抗拿扑净、颖壳色、抽穗期、穗轴、粒色、穗紧实度、抗逆性、穗重等;对数量性状的表型指标进行变异分析,其中株高的变异系数最大,为41.25%,抽穗期的变异系数最小,为11.03%;对20份矮秆突变体材料进行产量性状分析,发现E-13、E-58和E-98可作为矮秆、早熟、高产的材料;突变体中发现褐米色、抗拿扑净等优质、适宜产业化的材料。本研究构建的谷子突变体库具有丰富的突变类型,为谷子分子遗传学和育种提供了优异的基础材料。     

Molecular and morphological data reveals new insights into genetic diversity and population structure of Chinese cherry (<Emphasis Type="Italic">Prunus pseudocerasus</Emphasis> Lindl.) landraces

Chinese cherry (Prunus pseudocerasus Lindl.) is an indigenous fruit crop with high nutritional and ornamental value. It is widely cultivated in China, and many landraces have accumulated in large distribution regions. Genetic diversity analysis of Chinese cherry landraces is useful for breeding programs, as it helps to select genetic material to be used for further crossing. In this study, 110 Chinese cherry accessions from nine populations were assessed using fifteen morphological traits and seventeen ISSR markers. High average coefficient of variation (CV?=?17.43%) was detected based on morphological analysis. For molecular characterization, the global gene diversity (h?=?0.2816) and Shannon information index (I?=?0.4253) suggested a moderate high level of genetic diversity. Model-based STRUCTURE and principal coordinate analysis revealed three and two major gene pools based on morphologic and molecular data, respectively. Distinct distribution patterns of genetic variation between samples from southwest China and north China indicated two potential original domestication locations. We inferred that botanical features such as predominantly selfing characteristics, long lifespan, and insects-pollinated trait lead to the survival of abundant genetic variation and special genetic structure. This study provided morphological and molecular evidences for understanding genetic diversity and new insights into genetic consequence of populations for Chinese cherry landraces. Important implications for breeding programs and resources conservation were also provided.     

Analysis of average standardized SSR allele size supports domestication of soybean along the Yellow River

Soybean (Glycine max) was domesticated in China from its wild progenitor G. soja. The geographic region of domestication is, however, not exactly known. Here we employed the directional evolution of SSR (microsatellite) repeats (which mutate preferentially into longer alleles) to analyze the domestication process and to infer the most ancestral soybean landraces. In this study, the average standardized SSR allele sizes across 42 SSR loci in 62 accessions of G. soja were determined, and compared with those in 1504 landraces of G. max, collected from all over China and representing the diversity in the gene bank. The standardized SSR allele size in the landraces (0.009) was significantly (P = 8.63 × 10^?58) larger than those in G. soja (?0.406). Pairwise comparisons between inferred clusters and sub-clusters of Chinese landraces indicated that the average standardized SSR allele size also increased with the further differentiation of landraces populations. Spring-sowed types had the shortest size, followed by summer-sown types, while the sub-cluster of autumn-sown type had the largest length. The spring-sowed landraces located near the middle region along the Yellow River had the smallest allele sizes, indicating that this is the most ancestral population of cultivated soybean. We concluded that soybean was most likely domesticated in the middle region of the Yellow River in central China, initially as a spring-sown type.     

Genetic diversity of melon landraces (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) in the Xinjiang Uygur Autonomous Region on the basis of simple sequence repeat markers

We investigated the genetic variation and relationships among 35 melon landraces collected from the Xinjiang Uygur Autonomous Region in northwestern China by using 19 polymorphic simple sequence repeat markers (SSRs). A total of 55 polymorphic alleles were amplified. The number of alleles per SSR locus ranged from 2 to 5 with an average of 2.89 alleles per locus. The average gene diversity (GD) was 0.42 with a range of 0.06–0.71, and the average observed heterozygosity was 0.22 with a range of 0.06–0.97, indicating that the genetic diversity among the Xinjiang melon landraces was abundant. Genetic variation was also detected between the landrace populations in different regions in Xinjiang. The most abundant genetic diversity was observed among the landraces in Eastern Xinjiang, with the highest GD of 0.45 and PIC of 0.39. Eleven alleles (20 %) were found exclusively in the landraces from Eastern Xinjiang, and two alleles (3.6 %) were unique to the landraces from Southern and Northern Xinjiang. The genetic similarity matrix was defined on the basis of Jaccard coefficient to determine the genetic relationships among Xinjiang landraces. Cluster analysis was performed using the unweighted pair group method with arithmetic means, showing that the ‘wild Hami’ (XJ-34) landrace was distinct from the 34 other landraces that were divided into three clusters. Therefore, the genetic background of XJ-34 differed from that of the other landraces. The landraces were not precisely separated on the basis of their geographic origins, although most of these landraces were likely grouped near one another, as visualized through principal coordinate analysis. Thus, western China is one of the primary or secondary centers of melon diversity because of the relatively higher genetic variation detected among Xinjiang landraces. Except the ‘wild Hami’ landrace, Xinjiang melon landraces could be classified into two botanical varieties, namely, var. inodorus and var. cantalupensis. However, the distinction between these two genotypes was not significantly different.     

Intraspecific and interspecific variation in Setaria revealed by RAPD analysis

Random amplified polymorphic DNA (RAPD) analysis was applied to foxtail millet (Setaria italica) and other Setaria species in order to assess the degree of polymorphism among and within the species and to investigate their genetic relationships. Twenty accessions of foxtail millet, five of green foxtail (S. viridis), five of yellow foxtail (S. glauca), one each of S. germanica, S. verticilliformis, S. pallidifusca, S. macrostachya, S. sphacelata and S. faberi, and one unidentified, were evaluated for variability using a set of 19 random 10-mer primers. A total of 148 markers were scored for all the accessions. Wide variations in banding profiles among species were observed with nearly every primer used. The results showed that foxtail millet and S. glauca had a considerable degree of variation. The geographic differentiation of RAPD patterns in foxtail millet accessions examined seemed to exist. The present study confirmed that foxtail millet is more closely related to green foxtail than to other species in Setaria, providing support for the origin of foxtail millet from S. viridis and the conspecificity of these two species. The results confirmed that S. germanica and S. italica are conspecific and the former can be replaced by the latter. Setaria glauca and S. sphacelata are distinct from foxtail millet, implying that elite traits found in these two species are relatively difficult to be used in foxtail millet improvement. This study demonstrates that RAPD analysis can be used in assessing intraspecific and interspecific variation in foxtail millet and its wild relatives.