普通菜豆基因组学及抗炭疽病遗传研究进展

普通菜豆是重要的食用豆类之一,在世界各大洲普遍种植。近年来,普通菜豆在遗传图谱构建、新标记开发与利用、抗性基因定位以及比较基因组学等方面取得了很大进展。遗传连锁图谱的构建是基因定位与克隆的基础,是遗传研究中的重要内容;利用分子连锁图谱鉴定、标记和定位抗病基因将在种质改良和分子标记辅助育种方面发挥重要作用。豆科植物比较基因组学的研究成果为菜豆遗传连锁图谱的发展提供了新的思路。本文从普通菜豆遗传连锁图谱的获得、普通菜豆与大豆同线性比较以及抗炭疽病基因定位等方面进行了综述,以期为普通菜豆遗传改良和抗病育种提供参考。 关键词：普通菜豆;遗传连锁图;同线性比较;抗菜豆炭疽病     

水杨酸诱导普通菜豆镰孢菌枯萎病抗病性的研究

普通菜豆是人类主要食用豆类之一,其营养价值高、栽培面积大。镰孢菌枯萎病是普通菜豆典型的土传病害,给普通菜豆生产带来严重损失。水杨酸(SA)被认为是诱导植物抗病反应的重要信号分子之一,参与植物的过敏反应(HR)和系统获得性抗性反应(SAR)。本研究通过不同植物激素处理普通菜豆BRB-130,结果表明,SA处理普通菜豆叶片使植株根中SA的含量升高,并显著提高植株对枯萎病原菌FOP-DM01菌株的抗性。SA诱导普通菜豆根组织中苯丙氨酸解氨酶、过氧化物酶活性及过氧化氢的含量显著升高,从而诱导普通菜豆产生HR和SAR。因此,SA作为普通菜豆抗病信号途径中重要的化学激活因子,能够显著提高普通菜豆对枯萎病原菌的抗病性,为发展环境友好型化学农药提供新的思路。     

褪黑素对盐胁迫下普通菜豆芽期核酸修复的调控机制

普通菜豆(Phaseolus vulgaris)是重要的食用豆作物, 然而其极易受盐胁迫危害, 导致产量下降。褪黑素能提高植物耐盐能力。为探明外源褪黑素调控普通菜豆耐盐能力的机制, 以普通菜豆品种奶花芸豆(GZ-YD014)为实验材料, 设置水(W, 对照)、盐胁迫(S)和盐胁迫+100 µmol∙L^-1褪黑素(M+S) 3个处理。结果发现, 盐胁迫抑制了普通菜豆胚根的生长, 使其长度、表面积、体积以及直径显著降低, 外源褪黑素可缓解盐胁迫对普通菜豆胚根生长的抑制。外施褪黑素显著降低盐胁迫下活性氧积累和丙二醛(MDA)含量, 提高保护酶(过氧化物酶、超氧化物歧化酶、过氧化氢酶以及抗坏血酸过氧化物酶)活性, 增加渗透调节物质(可溶性糖和可溶性蛋白)以及生长素(IAA)、赤霉素(GA)和玉米素(ZT)的含量, 降低脱落酸(ABA)含量。通过转录组分析挖掘出217个差异表达基因(DEGs), DEGs在GO富集中显著(P-value<0.05)富集到核酸相关条目上, 在KEGG富集中显著(P-value<0.05)富集到核酸损伤修复(包括碱基切除修复、错配修复以及核苷酸切除修复)通路。qRT-PCR以及RAPD分析结果表明, 核酸损伤修复通路为外源褪黑素调控普通菜豆耐盐能力的一种机制。该研究揭示了外源褪黑素对普通菜豆芽期耐盐能力的调控机制, 可为褪黑素应用于盐胁迫下普通菜豆增产提供理论依据。     

普通菜豆种质资源芽期抗旱性鉴定

摘要：干旱是影响我国普通菜豆生产的主要因素之一,筛选芽期抗旱性种质资源,培育抗旱品种,有利于提高普通菜豆品种的出苗率和幼苗长势,对发展我国普通菜豆生产具有重要意义。本研究首先以4份普通菜豆种质为材料,检测了不同渗透势PEG6000溶液模拟旱胁迫下的发芽率和发芽势,确定了PEG6000溶液的最适渗透势为-0.7MPa（浓度为19.6%）;以-0.7MPa的PEG6000溶液对121份普通菜豆种质进行芽期模拟旱胁迫,测定发芽率、发芽势、下胚轴长、胚根长、干重和鲜重等10项指标;通过主成分分析筛选出相对发芽率、相对发芽势、相对鲜重、相对干重、相对胚根长,相对总芽长,相对胚根/下胚轴指数、相对发芽指数、相对活力指数等9项指标可以有效评价普通菜豆的芽期抗旱性;利用隶属函数分析法对121份种质的芽期抗旱性进行综合评价,筛选出跃进豆（F0000156）、白扁豆（F0000613）等芽期抗旱性种质,为普通菜豆抗旱生理与机制研究、抗旱育种奠定了基础。     

中国普通菜豆种质资源朊蛋白变异及多样性分析

朊蛋白是研究普通菜豆遗传多样性的一种重要且有效的生化标记。本试验通过SDS-PAEG凝胶电泳检测中国普通菜豆种质资源的朊蛋白变异类型,分析中国普通菜豆种质资源的遗传多样性及组成特点。来自中国13个省(自治区)的445份供试材料共检测到S、Sb、Sd、B、C、CA、T、PA、To、H、H1、CH 12种朊蛋白类型,表明中国普通菜豆种质朊蛋白变异类型丰富,遗传多样性水平较高。其中,Sb型朊蛋白种质最多,占比29.0%;T型其次,占比28.1%。依据朊蛋白类型在不同基因库的特异性,将研究材料明显地区分为中美基因库和安第斯基因库两大类。研究还发现中国普通菜豆种质资源中地方种质朊蛋白类型变异丰富,多样性明显高于现代育成品种或品系。最后,对种质朊蛋白类型与百粒重、子粒颜色、粒型进行相关性分析,结果表明朊蛋白类型与百粒重呈极显著正相关,而朊蛋白类型与子粒颜色、粒型2个性状之间无明显相关性。本研究结果将为普通菜豆种质资源的保护及有效地挖掘优质种质资源提供理论依据。     

基于农艺性状的山西普通菜豆初级核心种质构建

利用663份山西省普通菜豆资源基于14个农艺性状,采用比较不同分组原则、取样比例和总体取样量不同组合的取样方法,确定了"地理来源+平方根比例+20%总体取样量"为山西省初级核心种质构建的方法。同时,在此基础上对663份资源中一些具有极端性状的资源进行选择,最终确定152份普通菜豆可作为山西省普通菜豆初级核心种质。通过总体与初级核心种质资源多样性分析,数量性状均值比较,数量性状极值、变幅和标准差比较,性状多样性的差异性分析和各性状总体分布的χ2检验,最终得出:152份普通菜豆资源能够代表山西省普通菜豆资源的总体,可作为山西省普通菜豆评价和创新利用的优先样品集。     

菜豆种质资源抗普通细菌性疫病鉴定

采用针刺叶片接种法、温室地面水层保湿方式,对146份普通菜豆种质资源进行抗普通细菌性疫病鉴定和评价,结果表明,该接种方法简便实用,保湿方式效果良好,鉴定结果准确;从146份普通菜豆种质中筛选出抗病种质(R)2份、中抗(MR)种质50份、感病(S)种质81份、高感种质(HS)13份。本研究表明针刺叶片接种、温室地面水层自然蒸发保持湿度可以作为大规模菜豆抗普通细菌性疫病鉴定的适宜方法。     

普通菜豆抗炭疽病地方品种的朊蛋白标记分析

普通菜豆在长期驯化过程中形成了安第斯和中美两个基因库,研究基因库来源对于抗病育种中抗病亲本的选配具有重要意义.本研究利用菜豆朊蛋白标记分析了54份抗炭疽病菜豆地方品种的基因库来源,基本上明确了我国抗炭疽病菜豆种质的基因源,为抗病育种奠定了基础.     

普通菜豆SSR分子标记鉴定体系的建立及应用

为了保护育种家合法权益、规范种业市场、提高种子市场监管效率等,本研究拟建立一套普通菜豆品种SSR分子标记鉴定体系。首先,遴选12个产地不同、植物学形态差异较大的普通菜豆品种,通过聚丙烯酰胺凝胶电泳筛选出120对多态性高、条带清晰的SSR引物;然后,利用荧光毛细管电泳技术对筛选出的引物多态性在96个普通菜豆品种中进一步检测,最终获得在菜豆基因组上均匀分布、多态性高、峰型稳定的33对SSR核心引物。在此基础上,依据扩增片段长度和荧光标记颜色对核心引物进行分组,并设置参照品种以校正不同实验批次和平台之间的误差,建立了基于SSR标记的品种鉴定体系。同时,构建了包含申请品种、保护品种、主栽品种和地方品系在内的161个普通菜豆品种的指纹数据库。这套核心引物能够将其中的140个品种区分开,遗传相似系数平均值为0.33,表明这些品种遗传多样性较丰富;未区分开的21个品种在产地、父母本、植物学性状等方面极其相似,但能通过部分性状加以区分,表明该套核心引物在辅助近似品种筛选方面非常有效。本研究建立的基于SSR分子标记的普通菜豆品种鉴定体系将为其真实性鉴定及DUS测试近似品种辅助筛选提供有力支撑。     

中国普通菜豆形态性状分析及分类

对129份中国普通菜豆地方品种的形态性状进行分析,结果表明,8个性状共检测到35个变异类型,平均变异类型为4.375个,平均多态信息含量为0.5638。中国普通菜豆包括安第斯和中美两个基因库种质,中美洲基因库资源在参试资源中比重较大,但安第斯基因库资源遗传多样性水平高于中美基因库材料。由中美基因库向安第斯基因库渗透的天然杂交种质可为普通菜豆高产、优质、抗逆育种提供有价值的桥梁品种。     

国家库贮藏20年以上种子生活力与田间出苗率监测

对34种作物1.4万多份的国家库贮存种子进行了生活力监测,结果表明,贮存20年后,92.9%被监测种子的发芽率仍保持在85%以上,但有155份种子（占被监测份数的1.1%）出现了明显的下降（发芽率从80%以上降至70%以下）。发芽率显著下降的作物包括蚕豆、红小豆、黄麻、蓖麻、甜菜、西瓜、烟草、牧草。小麦等8种作物2078份种子的田间出苗率调查表明,所有种质均有出苗,但有8份种质的出苗率低于10%。出苗率与入库初始平均发芽率存在差别,平均出苗率最高的作物为普通菜豆86.2%,比该批入库初始发芽率平均值低9.3%;平均出苗率最低作物为谷子39.2%,比该批种子的入库初始发芽率平均值低51.3%。总体而言,国家库内保存的作物种子中,多数可安全保存20年以上,尤其是水稻等禾谷类作物种子,但对于蚕豆、红小豆等平均监测发芽率出现显著下降的作物种子需要增加监测频率,以确保种子的长期安全保存。     

Genetic diversity of common bean (Phaseolus vulgaris L.) ecotypes from Pakistan using Simple Sequence Repeats

Common bean (Phaseolus vulgaris L.) is a legume crop grown all over the world and is a very important food of mountain population of Pakistan for protein intake. The Western Himalayan Mountains are rich in biodiversity including unexplored landraces of the common bean crop. Unfortunately, very little attention has been given to this valuable crop in Pakistan, and it is being exported, majorly from Ethiopia, to meet the country’s requirements. The exploitation, utilization, preservation and multiplication of existing germplasm within the area are very important for sustainable production of the crop and enhancing the nutrition value for the local community in mountain regions. A research study was conducted for evaluation of biological diversity of common bean landraces from Azad Kashmir and Northern areas of Pakistan using morpho-physiological and molecular markers. Thirty-five common bean ecotypes along with one check variety were collected from different altitudes of Azad Kashmir and Northern Pakistan and screened for biological diversity. Morphological characterization revealed high genetic diversity in parameters including stem anthocyanin, growth type, days to flowering, pods/plant and 100 seeds weight. Genomic characterization using SSR markers, for allelic diversity evaluation among germplasm, also provided diverse profile with 83.3% polymorphism in banding pattern. The bulk of gene pool diversity evaluated within bean landraces may help to initiate breeding program for common bean improvement.     

Inoculation with indigenous <Emphasis Type="Italic">rhizobium</Emphasis> strains increases yields of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) in northern Spain,although its efficiency is affected by the tillage system

Common bean (Phaseolus vulgaris L.) crops hold the potential to obtain higher yields by enhancing their biological nitrogen fixation (BNF) with Rhizobium. However in contrast to other legumes, common bean has shown a lack of positive response to inoculation with Rhizobium in many cases. This has led to a limited use of rhizobial inoculants in this crop, especially in Europe. The adaptation of bacterial strains to the rhizosphere is a key factor in the success of any inoculant, especially in a promiscuous legume such as common bean. This research aimed at increasing common bean yields via inoculation with effective indigenous Rhizobium leguminosarum strains. Three highly effective strains (LCS0306, LBM1123 and ZBM1008) which were selected according to their effectiveness at BNF in hydroponic conditions were separately inoculated onto common bean in a field experiment. The experiment was carried out under three environments and three tillage systems: conventional-tillage (CONVT), no-tillage (NT) and a cover-crop (CC). The grain yield observed with seed inoculation was significantly higher than the yield obtained with uninoculated seed under CONVT and CC. However, under NT inoculation had no effect. Furthermore, under CONVT and CC, inoculation with R. leguminosarum LCS0306 produced even higher yields than those obtained in nitrogen-fertilised or control plots. This is the first attempt to explain the inoculation performance of common bean under different tillage systems in Europe. A gene–based hypothesis has been used to explain the effectiveness of indigenous common bean rhizobia as nitrogen fixers in this crop.     

New gene-derived simple sequence repeat markers for common bean (Phaseolus vulgaris L.)

Common bean is an important and diverse crop legume with several wild relatives that are all part of the Phaseoleae tribe of tropical crop legumes. Sequence databases have been a good source of sequences to mine for simple sequence repeats (SSRs). The objective of this research was to evaluate 14 sequence collections from common bean for SSRs and to evaluate the diversity of the polymorphic microsatellites derived from these collections. SSRs were found in 10 of the GenBank sequence collections with an average of 11.3% of sequences containing microsatellite motifs. The most common motifs were based on tri- and dinucleotides. In a marker development programme, primers were designed for 125 microsatellites which were tested on a panel of 18 common bean genotypes. The markers were named as part of the bean microsatellite-database (BMd) series, and the average polymorphism information content was 0.404 for polymorphic markers and predicted well the genepool structure of common beans and the status of the wild and cultivated accessions that were included in the study. Therefore, the BMd series of microsatellites is useful for multiple studies of genetic relatedness and as anchor markers in future mapping of wide crosses in the species.     

小豆种质资源、育种及遗传研究进展

小豆（Vigna angularis（Willd）Ohwi＆Ohashi）起源于中国,是我国主要食用豆类作物。该文综述了小豆种质资源的收集与利用、品种选育概况及经典遗传学、现代分子遗传学等方面的研究进展,可为豇豆属作物的研究提供参考。     

N2Fixation Response to Drought in Common Bean (Phaseolus vulgarisL.)

Nitrogen fixation activity in common bean is generally thoughtto be low and sensitive to soil drying and, consequently, droughtcan have important negative effects on N accumulation and yieldpotential. The objectives of this research were to examine theresponse of N₂fixation to drought stress in common bean, andto test the hypothesis that drought sensitivity of N₂fixationin common bean is linked to ureide levels in the plants. Twoglasshouse experiments were conducted to compare the responsesof leaf transpiration and acetylene reduction activity (ARA)to soil water contents. ARA decrease during soil dehydrationwas found to lag behind the decline in transpiration. This indicatesthat ARA is relatively less sensitive to soil dehydration comparedto leaf gas exchange. Further, in comparing two cultivars therewas no consistent difference in the relative response of ARAand transpiration to soil drying. The ureide concentrationsmeasured in common bean plants were low, ranging from 0.1 to1.0 mmol l^-1in xylem sap exudates. Ureide concentrations inthe sap exudate varied significantly among the two genotypeseven though there was no difference in ARA response to drought.It was concluded that in common bean, the lower sensitivityof N₂fixation to drought compared to leaf gas exchange couldbe related to low ureide concentrations in petioles and xylemsap.Copyright 1998 Annals of Botany Company Phaseolus vulgaris,nitrogen fixation, drought stress, nodules, ureides.     

Arbuscular mycorrhizal fungi induced changes in rhizosphere,essential oil and mineral nutrients uptake in dill/common bean intercropping system

The fundamental aim of this research was to evaluate the effects of Funneliformis mosseae on nutrient uptake and distribution in solo cropping and intercropping of common bean Phaseolus vulgaris and dill Anethum graveolens in the field, using a factorial arrangement of treatments. The factors were seven cropping systems including (a) common bean sole cropping (40 plants m^?2), (b) dill sole cropping at different densities (25, 50 and 75 plants m^?2) and (c) the additive intercropping of dill/common bean (25/40, 50/40 and 75/40 plants m^?2), with (+AM) or without (?AM) arbuscular mycorrhiza colonisation. In all cropping systems, AM colonisation improved dry weight and P, K, Fe, Zn and Mn contents of dill plants. Cultivable bacterial community was also increased in the dill/common bean intercropping. Yield of essential oil in dill shoot was improved as a result of AM colonisation and intercropping.