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.     

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.     

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.     

Application of random amplified polymorphic DNA to study genetic diversity in Paspalum scrobiculatum L. (Kodo millet, Poaceae)

Summary Genetic diversity and patterns of geographic variation among collections of Paspalum scrobiculatum (kodo millet) and P. polystachyum were studied using molecular markers generated through the random amplified polymorphic DNA (RAPD) method. A high level of polymorphism in RAPD markers was observed among the individual accessions, demonstrating the high genetic diversity of the crop. The markers obtained from the RAPD method were analyzed with the cluster analysis, principal coordinates and minimum spanning tree methods. Three major groups were resolved, one representing the African accessions, and two for the Indian accessions. The accessions of the north African kodo millet and P. polystachyum (considered conspecific with P. scrobiculatum) were quite distinct. The Australian kodo millet showed higher affinity to the African types. The study demonstrated that the RAPD technique can be applied to resolving degrees and patterns of genetic variation at the population and species levels, identifying cultivars, and defining gene pools of this crop.     

Geographic distribution of Waxy gene SNPs and indels in foxtail millet, Setaria italica (L.) P. Beauv.

To elucidate diversity and evolution of the Waxy gene in foxtail millet, Setaria italica, we analyzed sequence polymorphism of Waxy gene in 83 foxtail millet landraces collected from various regions covering the entire geographical distribution of this millet in Europe and Asia. We found a unique geographic distribution pattern at the sequence level of gene haplotypes and also found a large diversity in East Asian landraces. We also found a higher degree of genetic polymorphism in a non-waxy phenotype than in other low amylose types, supporting the hypothesis that low amylose types recently originated from non-waxy type.     

Genotyping of the <Emphasis Type="Italic">SiDREB2</Emphasis> gene in worldwide foxtail millet (<Emphasis Type="Italic">Setaria italica</Emphasis> (L.) P. Beauv.) genetic resources with special attention to Indian landraces

We genotyped Setaria italica Dehydration-responsive element-binding 2 (SiDREB2) gene, which had been reported to be associated with dehydration stress tolerance, in 588 accessions of foxtail millet from various parts of Eurasia and other regions by a dCAPS marker. Of these, 480 accessions were genotyped in ribosomal DNA (rDNA), polyphenol oxidase (PPO) gene and Heading date 1 (HD1) gene in our previous studies, and 108 accessions from India were newly used in this study and genotyped in these genes in addition to the SiDREB2 gene. We compared the geographical distribution of the SiDREB2 genotypes with that of these three genes. Accessions from countries in South Asia including India and Sri Lanka were the most variable in the SiDREB2 gene followed by Korean accessions and Japanese accessions, suggesting that Indian accessions and East Asian accessions are useful as genetic resources for dehydration stress tolerance. This study also showed that although Indian accessions are not so diverse in rDNA and in transposon display markers previously studied, they are diverse in adaptive genes such as SiDREB2, PPO and HD1 genes.     

大气CO2浓度升高对谷子生长发育及玉米螟发生的影响

人类活动导致全球大气CO_2浓度持续升高,研究大气CO_2浓度升高对C4作物谷子(Setaria italica)生长发育及虫害发生的影响,可以为谷子等C4作物制订应对气候变化栽培措施提供理论依据。本研究利用OTC(Open Top Chamber)系统,设两个CO_2浓度梯度(正常大气CO_2浓度、正常CO_2浓度+200μmol·mol-1)模拟CO_2浓度升高对谷子生长发育的影响。结果表明:大气CO_2浓度升高后,谷子净光合速率(Pn)、气孔导度(gs)、叶片蒸腾速率(Tr)和水分利用率(WUE)分别增加38.73%、27.53%、6.93%和40.56%;谷子叶片光系统Ⅱ最大光化学量子产量(Fv/Fm)和非光化学淬灭系数(NPQ)显著下降,光系统Ⅱ实际光化学量子产量(ΦPSII)和表观电子传递效率(ETR)显著增加,而对光化学淬灭系数(q P)无显著影响;此外,谷子株高、茎粗和小穗数分别增加3.41%、13.28%和13.11%;而叶重、茎重、千粒重、单株粒数和产量无显著变化,穗重和地上部分生物量分别显著下降12.8%和7.44%;大气CO_2浓度升高后,谷子灌浆期和收获期玉米螟(Ostrinia furnacalis)发生数量显著增加。大气CO_2浓度升高将有利于谷子的生长发育,但会增加玉米螟危害。     

Why no tetraploid cultivar of foxtail millet?

Autotetraploid material was induced by colchicine treatment of a pure line of foxtail millet and an interspecific hybrid between foxtail millet and giant green foxtail. Compared with diploid material, tetraploids were smaller, flowered later, and had a two-fold reduced fertility (number of grain per cm of spike). Grain weight increased by 20% with polyploidy, but total grain yield decreased by 46%. This work illustrates the difficulty of breeding polyploid lines of foxtail millet that are of agronomic use.     

Assessment of genetic diversity in Solanum trilobatum L., an important medicinal plant from South India using RAPD and ISSR markers

Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.     

Assessment of genetic diversity and genetic relationships among 29 populations of <Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss. using RAPD markers

Randomly amplified polymorphic DNA (RAPD) analysis was employed to assess genetic divergence among 29 neem accessions collected from two agro-ecological regions of India (11 agro-climatic sub-zones), which cover three states, Punjab, Haryana and Rajasthan. Out of 24, 10-mer random primers used for studying genetic divergence, 14 were polymorphic, generating a total of 73 amplification products with an average of 5.21 products per polymorphic primer and estimated gene diversity of 0.49. Genetic relationships among accessions were evaluated by generating a similarity matrix based on Jaccard’s coefficient, ranging from 0.70 to 0.96. The phenetic dendrogram generated by UPGMA analysis grouped accessions into five clusters. RAPD performed within accessions (individual seedlings collected from the same mother plant) showed no variation indicating homogeneous population within accessions. Primers OPA-18, OPC-08 and OPI-03 were found most informative based on their resolving power. The degree of genetic variation detected among the 29 accessions with RAPD analysis suggests that RAPD can be used for studying genetic diversity in neem. The study also demonstrated that neem germplasm collected from northwestern plains of India shows no eco-geographical isolation based on sub-zones because accessions collected from different sub-regions are grouping together in the genetic tree.     

谷子播期对谷子/花生间作系统生产力的影响

间作作物的播期会影响其共生期,进而影响作物生产力和种间相互作用。为探究谷子(Setaria italic L. Beauv)播期对谷子/花生(Arachis hypogaea L.)间作系统生产力的影响,以谷子金选6号和花生大花生606为试验材料,固定花生播期为5月15日,谷子从5月15日至6月9日每5天播种1期,共设置6个播期处理(S1~S6)。通过对单间作系统中作物产量、产量构成因素等指标的测定,分析不同谷子播期条件下谷子/花生间作的作物产量、系统生产力、种间竞争力的变化。结果表明,随着谷子播期的推迟,间作系统中谷子和花生的产量均表现为先升高后降低的趋势,5月25日播种时(S3)谷子产量(4.07 t·hm^-2)和土地当量比(LER=1.24)最大,5月30日播种时(S4)花生产量(2.51 t·hm^-2)、间作总产值(4.64万元·hm^-2)和间作系统生产力(SP=2.93 t·hm^-2)最大;谷子/花生间作系统的实际产量损失大于0,具有产量优势。谷子的侵占力大于0、竞争比率大于1,花生的侵占力小于0、竞争比率小于1,间作系统中的谷子是竞争优势作物,对资源的竞争能力高于花生且受谷子播期影响较大。相关性分析及通径分析发现,在间作系统中,谷子产量与穗长呈显著正相关,与穗粗、穗重、穗粒重呈正相关,且穗长对谷子产量的贡献作用最大。本研究结果对谷子/花生间作实践具有指导意义。     

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。     

Assessment of Genetic Diversity among Finger Millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.) Accessions using Molecular Markers

Finger millet (Eleusine coracana), an allotetraploid cereal, is widely cultivated in the arid and semiarid regions of the world. Being rich in protein and calcium, finger millet serves as an important staple food for rural populations in developing tropical countries where calcium deficiency and anemia are wide spread. Thirty-two finger millet genotypes were fingerprinted using 50 random amplified polymorphic DNA (RAPD) markers. Out of the total 529 loci generated using the 50 RAPD primers, 479 loci (91%) were polymorphic and informative to differentiate the accessions. Cluster analysis grouped the 32 finger millet accessions into two major clusters. Among the 32 finger millet genotypes, GEC 182 and CO 12 were distantly related with a low similarity index of 0.315. These two accessions also differed considerably in days to flowering and grain weight; GEC 182 is early flowering and has bold grains, while CO 12 is late flowering and has smaller grains. These two accessions with higher diversity at molecular level, phenology and grain weight will be ideal as parents in hybridization programme, to develop improved finger millet varieties suitable for peninsular region of India.     

谷子抗除草剂基因的发现及其应用

谷子田间除草一直是阻碍谷子产业化发展的主要因素之一。谷子抗除草剂品种的出现,解决了谷子田间除草难的问题,有利于谷子大面积的栽培,使谷子产业化发展成为可能。本文围绕谷子抗除草剂基因的发现、类型和机理以及抗除草剂品种在谷子生产中的应用情况等方面进行阐述,探讨谷子抗除草剂应用对环境的影响,并对谷子抗除草剂研究中存在的问题及应用前景进行了展望。     

小麦田杂草种类及危害情况调查

调查掌握小麦田杂草种类、分布和危害程度,可为筛选高效除草药剂提供科学依据。对定西市不同区域的小麦田杂草发生情况开展了调查,结果表明,定西市共有麦田杂草15科38种,其中单子叶杂草6种,双子叶杂草32种。安定区、通渭县、陇西县、临洮县以及渭源县北部等干旱半干旱区分布较多的杂草有12种,渭源县南部以及漳县、岷县等二阴区分布较多的杂草有13种。有8个杂草优势种,其中苦苣菜、藜、狗尾草、灰绿藜、菊叶香藜等5种杂草危害较重,危害等级达到Ⅲ级;苦苣菜、藜、狗尾草为绝对优势种,危害最重。杂草优势种是防除的重点对象,建议结合杂草特性,筛选高效除草剂加以科学防除。     

花青素积累与谷子低温胁迫响应的关系

为探究花青素积累与谷子低温胁迫响应的关系,本研究以绿叶和富含花青素的紫叶谷子品种为材料,通过对不同品种幼苗进行低温和常温(对照)处理,观察叶片中色素分布,测定叶片光合作用以及花青素、相对叶绿素、丙二醛(MDA)、脯氨酸(Pro)、可溶性糖(SS)含量。结果表明,低温诱导后紫叶品种叶片花青素主要分布在叶肉细胞和上下表皮细胞,而绿叶品种叶片中几乎无花青素积累。低温诱导后紫叶品种花青素变化明显,绿叶品种无明显变化;与对照相比,低温诱导12 d紫叶品种B13、B45、B55花青素含量分别增加了234%、226%、126%。低温诱导后,紫叶品种叶片的净光合速率(Pn)和相对叶绿素含量均降低,但SS和Pro含量显著升高,MDA含量无规律性变化。相关性分析结果表明,低温诱导12 d紫叶品种叶片花青素含量与相对叶绿素含量、Pn呈极显著负相关,而与Pro含量、SS含量呈极显著正相关,相对叶绿素含量与Pn呈正相关。由此可知,紫色谷子品种叶片花青素的合成可以被低温诱导,且与Pro、SS共同参与植物对低温胁迫的响应。本研究为探索紫叶谷子的抗寒性以及扩大其种植区域提供了理论依据。     

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。     

RAPD variation in wild, weedy and cultivated azuki beans in Asia

RAPD (Random Amplified Polymorphic DNA) variation was assessed in 42 accessions of azuki bean (Vigna angularis) including wild, weedy and cultivated races and in three accessions of two related species used as outgroups. A much lower level of genetic variation was observed in cultivated and weedy azuki beans compared to wild azuki bean. Wild azuki bean (V. angularis var. nipponensis) has relatively high genetic variation in subtropical highlands of Asia compared to the Far East. Although cultivated azuki bean has low RAPD variation, accessions from subtropical highlands and Southeast Asia showed different RAPD features compared to those of the Far East. It is hypothesized that the cultivated azuki bean has been derived from wild azuki bean in the Far East; the high variation in wild azuki bean has been created through its natural dissemination; and the relatively low variation in cultivated azuki bean has come about through human dissemination after genetic bottleneck reduced by domestication. In addition, high genetic diversity in wild azuki bean in subtropical highlands of Asia is regarded as an important genetic resource in azuki improvement.     

Genetic diversity in cowpea [Vigna unguiculata (L.) Walp.] as revealed by RAPD markers

The present study, using RAPD analysis, was undertaken to characterize genetic variation in domesticated cowpea and its wild progenitor, as well as their relationships. The materials used consisted of 26 domesticated accessions, including accessions from each of the five cultivar-group, and 30 wild/weedy accessions, including accessions from West, East and southern Africa. A total of 28 primers generated 202 RAPD bands. One hundred and eight bands were polymorphic among the domesticated compared to 181 among wild/weedy cowpea accessions. Wild accessions were more diverse in East Africa, which is the likely area of origin of V. unguiculata var. spontanea. Var. spontanea is supposed to have spread westward and southward, with a loss of variability, loss counterbalanceed in southern Africa by introgressions with local perennial subspecies. Although the variabilty of domesticated cowpea was the highest ever recorded, cultivar-groups were poorly resolved, and several results obtained with isozyme data were not confirmed here. However primitive cultivars were more diverse than evolved cultivars, which still suggests two consecutive bottlenecks within domesticated cowpea evolution. As isozymes and AFLP markers, although with a larger number of markers, RAPD data confirmed the single domestication hypothesis, the gap between wild and domesticated cowpea, and the widespread introgression phenomena between wild and domesticated cowpea.