Inheritance characteristics of F1 and F2 offsprings obtained from full diallele crosses of five cowpea cultivars (Vigna unguiculata L. Walp) against pseudocercospora leaf spot disease

The experiment was carried out at the Experimental Farm, Department of Agricultural Technology, Mahasarakham University, Mahasarakham Province in the 2005 (February to April) for F1 offsprings and also at the Experimental Farm, Khon Kaen University, Khon Kaen Province, Northeast Thailand for F2 offsprings (July to October), i.e., four experiments were carried out, each location had two experiments. The research aimed to investigate inheritance characteristics of F1 and F2 offsprings derived from full diallele crosses of five cowpea cultivars (20 pairs of crosses) against a pathogenic disease of Pseudocercospora cruenta (Sacc,) Deighton. The first two experiments were carried out under field and glasshouse conditions at Mahasarakham University for F1 offsprings and the other two experiments were carried out at Khon Kaen University for F2 offsprings. A Chi square test method was used to justify dominant genes on the infection of the disease. The results showed that the cowpea plants of F1 and F2 offsprings gave only one pair of dominant gene that had its complete dominant effect over recessive gene of the cowpea offsprings.     

Soil nutrients and liming on dry weight yields and forage quality of Signal grass (Brachiaria decumbens Stapf.), grown on Korat soil series (oxic paleustults) in northeast Thailand

This experiment was carried out at Khon Kaen University Experimental Farm, Khon Kaen University, Thailand during the 2004-2005 aiming to investigate effect of phosphorus (P) and dolomite levels on dry weight yields (DWYs) and forage quality of Signal grass. A 4x3 factorial arranged in a Randomized Complete Block Design (RCBD) was used. Four P levels were: 0, 100, 200 and 400 kg P2O5 ha-1 and three dolomite levels were: 0, 625 and 2,500 kg ha-1. The Signal grass plants were grown on Korat soil series, (Oxic Paleustults). A quadrat with a dimension of 50x50 cm was used for grass yield harvests. Crude Protein (CP), Acid Detergent Fibre (ADF), Neutral Detergent Fibre (NDF) and Dry Matter Degradability (DMD) contents were determined. Tissues phosphorus and calcium contents were also analysed. The results showed that an increase in dolomite levels increased soil pH from 4.4 to 5.1 for levels 1 and 3, respectively. An increase in P levels increased available soil P from 4.56 to 28.38 ppm for levels 1 and 4, respectively. For the first year experiment, dolomite levels had no significant effect on DWYs, whilst P levels significantly increased but only up to level 2. The 2-year average DWYs reached 11,368 kg ha-1 for level 4 of P. With the first year rainy season harvests, P levels had its significant effect on ADF and DMD up to level 2 but not with CP and NDF. For the dry season harvests, P and dolomite levels had no significant effects on forage quality. Dolomite levels had no significant effect on P and Ca contents of the Signal grass tissues but an increase in P levels increased P contents. P and Ca contents, in most cases, were higher for the dry season than the rainy season.     

利用SSR分子标记研究花生属种间亲缘关系

以5份花生栽培种资源和花生属六个区组的19份野生种资源为研究材料，通过SSR分子标记技术分析其DNA多态性并进行聚类分析。大多数从栽培种基因组分离出的SSR引物能在野生种中扩增出DNA片段，共筛选出21对多态性SSR引物；每对引物在花生基因组中扩增出的条带数为1～13条，在供试材料中扩增出的总条带数为5～40条，平均18.1条，其中多态性条带为4～40条，平均18.0条；SSR引物的多态性指数为0.92～9.04；供试材料间的遗传距离为0.33～0.91，平均0.76。结果表明，大多数花生SSR引物为多位点引物，花生属种间种质存在丰富的DNA多态性， A. duranensis是花生栽培种（A. hypogaea L.）的野生种亲本之一，与花生区组野生种亲缘关系最近的是异形花区组，最远的是大根区组。     

墨兰"达摩"芽变叶艺品种亲缘关系的RAPD分析

利用RAPD分子标记对10个墨兰叶艺品种进行亲缘关系分析。从100个10碱基随机引物中筛选出17个多态性高的引物,共扩增出159条DNA带,其中137条(86.2%)为多态带,平均每个引物扩增DNA带数9.3条。10个叶艺品种之间的Dice相似系数变化范围为0.598-0.813,平均相似系数为0.706。RAPD扩增结果的UPGMA聚类分析的结果表明,墨兰叶艺品种间的亲缘关系与形态学分类结果基本一致,而且还与品种来源地密切相关。     

An evaluation on intensity of infection of Pseudocercospora leaf spot disease of cowpea cultivars (Vigna unguiculata L. Walp), with respect to infector rows, dates of inoculation and cultivars, grown under field conditions in Northeast Thailand

This investigation was carried out at the Experimental Farm, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand to evaluate intensity of infection of Pseudocercospora Leaf Spot disease of cowpea cultivars, grown under field conditions with the use of Yasothon soil series (Oxic Paleustults). The experimental design used was a strip split plot design with four replications. The experiment consisted of three factors, i.e., with and without infector rows (Factor A), with and without inoculation and inoculated dates, i.e. none inoculation, inoculated at days 15 and 30 after sowing of seeds in main plots (Factor B) and three cowpea cultivars of KVC7, KKU25 and IT81D-1228-14-1 (Factor C). The results showed that the use of infector rows of cowpea plants (KKU25) being sown at 15 days before sowing seeds of cowpea cultivars in main plots gave a similar intensity of disease as those infector rows being sown at 30 days before sowing seeds of cowpea cultivars in main plots but significantly higher than without infector rows. Pathogenic disease being inoculated to the cowpea plants at 15 days after the sowing of seeds in main plots gave significantly higher intensity of disease than none inoculation. The establishment of infector rows at 15 days ahead of the sowing of seeds in main plots together with an inoculation of the disease at 15 days after the sowing of seeds in main plots gave the best results in evaluating amount of the infected disease. Thus infector rows surrounded the main plots are required whenever trials on disease evaluation are taken place.     

Rooting traits of peanut genotypes with different yield responses to pre-flowering drought stress

Water stress during the vegetative development normally is not detrimental and sometimes actually increases yield of peanut (Arachis hypogaea L.). Root growth might play an important role in response to early season drought in peanut and might result in an increase in yield. Information on the response of root characters of diverse peanut genotypes to these conditions will provide useful information for explaining mechanisms and improving peanut genotypes for exploiting positive interaction for pod yield under pre-flowering drought. The aim of this study, therefore, was to investigate the root dry weight and root length density of peanut genotypes with different yield responses to pre-flowering drought stress and their relationships with pod yield. Field experiments were conducted at the Field Crop Research Station of Khon Kaen University, Khon Kaen, Thailand during February to July 2007 and during February to July 2009. A split-plot experiment in a randomized complete block design was used. Two water management treatments were assigned as the main plots, i.e. field capacity and pre-flowering stress, and six peanut genotypes as the sub-plots. Total crop dry matter, root dry weight and root length density were recorded at 25 DAE, R5 and R7. Top dry weight and pod yield were measured at harvest and pod harvest index (PHI) was computed using the data on pod yield and biomass. Peanut genotypes were categorized into three groups based on their responses to drought for pod yield, e.g. increasing, decreasing and non-responsive groups. The genotypes of each group showed a differential response for root quantity and distribution. The increasing pod yield group had more root dry weight and root length density in the deeper soil layers during pre-flowering stress compared to the non-stress treatment. The non-responsive group showed no root response under pre-flowering drought conditions compared to the non-stress treatment. A larger root system alone without considering distribution may not contribute much to pod yield but a higher RLD at deeper layers may allow plants to mine more available water in the sub-soil. However, as yield is a complex trait, several mechanisms may be involved. The increasing pod yield group also had the ability to maintain a high PHI.     

利用RAPD进行茶组植物遗传多样性和分子系统学分析

对张宏达系统山茶属茶组植物24种、变种的遗传多样性和分子系统学进行了RAPD分析。从冬梢中获得了高质量和高得率的基因组DNA, A260/A280平均为1.71,平均得率为331μg/g鲜重。从Operon技术公司的61个十聚体随机引物中筛选出15个用于茶组植物的RAPD扩增,在所扩增得到的107条可重现谱带中(平均7.1条/引物)有102条(平均6.8条/引物)是多态的,多态性程度是95.3%。每个引物扩增的谱带数在211条之间,相对多态性频度在0.04—0.96之间,不过总平均多态性频度只有0.30,     

玉米双低频酶cDNA-AFLP体系的建立

以玉米自交系QCL1094为材料,用RNAiso Reagent、柱式试剂盒法、异硫氰酸胍法等3种方法提取苗期叶片总RNA,用PrimeScriptTM 1st Strand cDNA Synthesis Kit反转录获得第一链cDNA,用RNase H、E.coli DNA聚合酶I和E.coli DNA连接酶合成第二链cDNA,用T4 DNA聚合酶进行末端平滑得到双链cDNA,最后,用双低频酶EcoR I和Pst I酶切双链cDNA,用T4 DNA连接酶连接接头,用引物EA00和P00进行预扩增,用引物EA01和P01进行选择性扩增。经1%琼脂糖凝胶和6%变性聚丙烯酰胺凝胶电泳检测,结果表明,RNA提取及cDNA-AFLP预扩增和选择性扩增的效果良好。     

新型植物油莎豆DNA提取与SRAP体系优化

以14个不同地理来源的油莎豆品系为实验材料,研究了油莎豆DNA快速提取方法;以SRAP反应体系中DNA模板量、Mg2+浓度和引物浓度3个因子分别设置3个水平,共配制27个反应体系,对油莎豆SRAP反应体系进行优化。研究结果表明:改良CTAB法可获得较高质量的DNA,参试材料的A260/A280介于1.70～1.98;在27个SRAP反应体系(15μL)中,最优反应体系为DNA模板25ng、Mg2+1.5mmol/L、引物浓度1μmol/L、dNTPs 0.3mmol/L和Taq酶1U,可扩增出清晰稳定的多态性条带。     

Association between aflatoxin contamination and drought tolerance traits in peanut

The current study investigates the association between drought tolerance traits and aflatoxin contamination in peanut grown under long-term drought. Two field experiments were conducted at Khon Kaen University, Thailand using a split–split plot design with three drought stress levels as main plots, 11 genotypes as sub-plots, and two soil inoculations of Aspergillus flavus treatments as sub-sub-plots. The effects of temperature, soil moisture and A. flavus population on kernel colonization and aflatoxin contamination, and drought tolerance traits viz. specific leaf area (SLA) and root length density (RLD) were measured. The results demonstrated that elevated soil temperatures and reduced soil moisture, favored aflatoxin production. Drought in combination with higher levels of A. flavus inoculum load in the soil resulted in an increase in the fungal populations in the soil which in turn resulted in increased kernel colonization and subsequent aflatoxin contamination. A combination of SLA and RLD, and kernel colonization had a significant influence on aflatoxin contamination under drought conditions in both seasons (r = 0.73** and 0.76**). The results revealed that drought tolerance traits (SLA and RLD) could be contributing to resistance to aflatoxin contamination suggesting that a combination of SLA, RLD and kernel colonization could be used as selection criteria in selecting parents for aflatoxin resistance.     

豆豉纤溶酶基因的克隆及其在枯草杆菌中的表达

以实验室筛选的UD8豆豉芽孢杆菌的总DNA为模板,应用PCR技术根据豆豉纤溶酶基因DNA序列(GeneBank AY720895.2)设计并合成1对引物,扩增出了豆豉纤溶酶基因,将该基因克隆到大肠杆菌-枯草芽孢杆菌穿梭质粒EC-BS8上,构建成表达质粒EC-BS8,再将其转化到枯草芽孢杆菌Bs6中。筛选表达菌株,实现了豆豉纤溶酶基因在枯草杆菌中高效表达。获得重组菌纤溶酶活力高达9 830 IU.mL-1,是出发菌株活力的2.9倍。经纯化SDS-PAGE鉴定重组豆豉纤溶酶相对分子质量为31.0 kDa。     

PCR Amplification of the Hypervariable Region of Wheat Triticin Genes

A modified extraction procedure for wheat triticin allowed a better resolution of the triticin bands on SDS-PAGE. The modified protocol was used for the verification of Chinese Spring ditelocentric stocks before DNA extraction for PCR amplification of triticin gene segments. Three pairs of PCR primers were designed for specific amplification of two overlapping segments of the triticin gene including its hypervariable region. Consistent amplification was achieved with only one pair of primers for which the PCR conditions were further optimised. The bulk of the amplified product was due to genes on the short arm of chromosome 1D. Hence, under the given conditions there was a preferential amplification of the Tri-D1 gene. These primers will be useful in screening for natural variation in the triticin HVR segment in wild wheat collections.     

稻瘟病菌生理小种RAPD分析及其与马唐瘟的差异

氯化卞法微量提取稻瘟病菌７个生理小各和马唐瘟病菌基因组ＤＮＡ，用６０个随机引物进行ＰＣＲ扩增，其中２１个引物的特异性扩增条带，第一引物－菌系组合扩增到３－１９条，平均１０．３条；其中有多态性条速为１－１２条，平均６．１条，多态性为６１．１％。说明稻瘟病菌群体有丰富的ＲＡＰＤ多态性。引物ＯＰＨ１、ＯＰＨ２，ＯＰＨ４，ＯＰＨ１３，ＯＰＨ１４，ＯＰＫ１４等扩增的条带多，多态性丰富，而且可以有效地区分稻瘟     

Evaluation of genetic diversity in turmeric (Curcuma longa L.) using RAPD and ISSR markers

Turmeric (Curcuma longa L.) is an industrially important plant used for production of curcumin, oleoresin and essential oil. In the present study we examined the genetic diversity among turmeric accessions from 10 different agro-climatic regions comprising 5 cultivars and 55 accessions. Two DNA-based molecular marker techniques, viz., random amplified polymorphism DNA (RAPD) and inter simple sequence repeat (ISSR) were used to assess the genetic diversity in turmeric genotypes. A total of 17 polymorphic primers (11 RAPDs and 6 ISSRs) were used in this study. RAPD analysis of 60 genotypes yielded 94 fragments of which 75 were polymorphic with an average of 6.83 polymorphic fragments per primer. Number of amplified fragments with RAPD primers ranged from 3 to 13 with the size of amplicons ranging from 230 to 3000 bp in size. The polymorphism ranged from 45 to 100 with an average of 91.4%. The 6 ISSR primers produced 66 bands across 60 genotypes of which 52 were polymorphic with an average of 8.6 polymorphic fragments per primer. The number of amplified bands varied from 1 to 14 with size of amplicons ranging from 200 to 2000 bp. The percentage of polymorphism using ISSR primers ranged from 83 to 100 with an average of 95.4%. Nei's dendrogram for 60 samples using both RAPD and ISSR markers demonstrated an extent of 62% correlation between the genetic similarity and geographical location. The result of Nei's genetic diversity (H) generated from the POP gene analysis shows relatively low genetic diversity in turmeric accessions of South eastern ghat (P7), Western undulating zone (P8) with 0.181 and 0.199 value whereas highest genetic diversity (0.257) has been observed in Western central table land (P9). Knowledge on the genetic diversity of turmeric from different agro-climatic regions can be used to future breeding programs for increased curcumin, oleoresin and essential oil production to meet the ever-increasing demand of turmeric for industrial and pharmaceutical uses.     

利用RAPD、ISSR分子标记分析野牡丹属亲缘关系

采用ISSR和RAPD分子标记技术对9个种44份野牡丹属种质资源进行了亲缘关系分析。结果表明:11条ISSR引物共扩增出91条谱带,其中多态性条带90条,多态性条带的比率为98.9%;16条RAPD引物共扩增出113条谱带,其中101条多态性条带,多态性比率为89.38%。2种分子标记相比较,ISSR分子标记检测出的多态性比率更高。44份野牡丹属种质明显聚为2组,种质资源遗传相似系数在0.61~0.88,平均相似系数0.75。基于不同引物的条带组合构建了供试的44份野牡丹属种质资源的ISSR和RAPD指纹图谱,采用这2种指纹图谱可对供试的所有野牡丹属种质资源进行鉴定。     

一个鉴别低温刺激型秀珍菇的ISSR-SCAR标记建立与应用

为了快速、准确地鉴别低温刺激型秀珍菇菌株,在对18个供试菌株进行拮抗试验、现蕾出菇试验、ISSR图谱分析的基础上获得了秀珍菇菌株的ISSR特异性标记,将其克隆、测序。根据测序结果,应用Primer 6.0软件设计SCAR引物进行引物筛选,获得1对引物并成功转化为低温刺激型秀珍菇的稳定SCAR标记。采用该引物对供试秀珍菇菌株的基因组DNA 进行PCR扩增,低温刺激型秀珍菇菌株均能扩增出大小为205 bp的特异性DNA片段。该ISSR-SCAR标记能快速、有效地鉴别出低温刺激型秀珍菇菌株,准确率达100%,为秀珍菇种质资源的鉴别和分类提供了技术支撑。     

大豆疫霉菌遗传多样性的RAPD分析

利用随机扩增DNA (RAPD)技术分析了15个大豆疫霉菌( Phytophthora sojae)的遗传多样性,并同其它7 个疫霉种的8个菌株进行了比较。运用筛选出的12个引物共获得152个RAPD标记,其中84. 2%具有多态性。通 过聚类分析结果表明,疫霉属种间的遗传相似系数的变化幅度为0. 533～0. 783,大豆疫霉菌与其它几种疫霉的遗 传距离相对较远,用引物OPB06扩增到P. sojae种特有的一个RAPD标记; P. sojae种内菌株间遗传相似系数的变化 幅度为0. 855～0. 967,亦存在较丰富的遗传多样性,该遗传多样性与菌株毒力之间有一定相关性,而与菌株地理来 源没有相关性。     

基于转录组测序的油梨 EST-SSR 引物开发

利用转录测序技术,开发油梨表达序列标签-简单重复序列（EST-SSRs）,为 SSR 标记在油梨种质资源鉴定、 品种选育及遗传连锁图谱构建奠定基础。采用 Illumina 二代测序的技术,共获得 37 639 条无冗余的序列,对其进行 SSR 搜索,共获得 6 419 条简单序列重复（SSR）。利用 Primer 3.0 软件设计 SSR 引物,并以 11 份油梨种质筛选多态性引物。 基于转录组序列开发出的 EST-SSR 的分布频率为 17.05%。在油梨 EST-SSR 中,单核苷、二核苷和三核苷的重复占主 导,占总数的 99.07%。单、二、三核苷酸重复单元分别占总 SSR 的 37.47%、31.80%和 29.80%;出现频率最高的二核 苷酸重复基元是 AG/CT,占总数的 29.15%,出现最高的三核苷酸重复基元为 AAG/CTT,占 12.01%。随机选择 315 个 SSR 位点合成引物,经 11 份油梨种质筛选鉴定,227 对引物可扩增获得产物,有效扩增率为 72.06%;其中 34 对引物 表现出良好多态性,占有效引物的 12.78%,占总引物的 10.79%。在 34 对多态性引物中,每对引物扩增等位基因数 2~12 个。利用高通量测序开发 SSR 引物有较好的实用性,开发获得的 34 个具有多态性的油梨 SSR 标记可用于研究油梨及 其相关近缘物种的遗传变异。     

Genetic variation of wild rice populations from Thailand and the Lao PDR based on molecular markers

The two wild rice species, Oryza rufipogon and O. nivara, which inhabit Thailand and the Lao PDR, are threatened with the loss of their natural habitats. The losses are primarily attributable to human intervention. Before this process advances, it is crucial to obtain basic information on the genetic variations of these species, so important knowledge about the current status of genetic variability. In this study, genetic variation within and between fourteen natural populations of the two populations from Thailand and Lao PDR was investigated at the DNA level by analysis of Polymerase Chain Reaction (PCR) mediate molecular markers. The results illustrated that wild rice from Thailand carried deletion (D) type ORF 100, but deletion and non-deletion type (ND) were found in wild rice samples from the Lao PDR. Five different plastid subtypes (7C7A, 6C8A, 7C6A, 6C7A and 9C7A) were found in the collected samples. Both polymorphism and pattern of distribution of p-SINE1-r2 in the two wild rice species were found among the populations. These data indicated that genetic variation existed in these natural populations of wild rice, suggesting that the strategies should be developed that are conducive to the conservation of wild rice in its natural environment.     

大豆抗旱种质资源遗传多样性的SSR分析

以45份具有不同程度抗旱性的大豆为材料,利用SSR分子标记对这些材料进行分析,选取2个产地、农艺性状差异显著的大豆品种静乐黑滚豆和辽13,对SSR引物进行筛选,从中选择出多态性丰富,带型较好的引物11对.用筛选出的引物对45份大豆种质进行PCR扩增.聚类结果表明:GD=0.41处可将45个大豆品种分成两大类,大部分品种被归为第1类,有31个品种,占总品种数的69%,第2类包含14品种,占总品种数的31%.两个大类又各自细分成不同的亚类群,聚类结果反映出品种间关系与地理起源、表型形态等具有一定的相关性.