与茄子青枯病抗性相关基因连锁的AFLP标记研究

本文以茄子高感青枯病亲本自交系5810、高抗亲本自交系5556单1、两自交系杂交得到F1,F1单株自交得到F2群体为试材,采用苗期接种鉴定及BSA法结合AFLP技术,探讨了茄子青枯病抗性的遗传规律,获得与茄子青枯病抗性相关基因连锁的AFLP分子标记。结果表明：抗病亲本5556单1青枯病抗性受一对单隐性基因控制,感病亲本控制的感病基因是不完全显性的。在抗、感池中未发现标记,而通过抗、感池单株分析得到了相引相AFLP分子标记E13M10,如及相斥相AFLP标记E16M5240,估算它们与目标基因间的遗传距离分别为10．14cM和7．56cM。     

玉米矮花叶病毒抗性资源鉴定的研究

利用人工接毒方法对 70份玉米种质资源进行了两年玉米矮花叶病毒B株系的抗性鉴定。依据病情指数 ( % )将抗病程度分为高抗、抗、中抗及感病 4个等级。试验筛选出高抗自交系 4份、高抗单交种 3个、抗病毒自交系 10份、抗病单交种 3个 ;中抗自交系 6份、中抗群体 3个。讨论了这些种质资源在我国抗玉米矮花叶病遗传及育种研究上的应用价值     

广西玉米自交系纹枯病抗病性鉴定

2003年秋至2005年夏连续2年对广西223份玉米自交系进行多年多重复的田间纹枯病抗病性鉴定。接种鉴定方法:将玉米纹枯病分离菌在大麦粒上扩繁,于玉米拔节期进行田间接种,采用国际玉米小麦改良中心(CIMMYT)的病害分级评定方法进行鉴定。结果表明:玉米自交系间的抗感性存在差异,同一自交系在同年度内春播与秋播鉴定中抗病性表现存在明显差异。在223份材料中未发现免疫类型,高抗自交系占所鉴定总数的1·3%、抗病的占14·4%、中抗水平占6·3%、感病占2·2%、高感占75·8%。     

利用phi116和umc1044标记选育抗纹枯病玉米品系

玉米纹枯病已成为我国玉米产区的主要病害之一,为解决玉米纹枯病抗病育种工作中人工接种鉴定困难、选择效率低等问题,利用第7和第1染色体上与抗病QTL紧密连锁的两个标记phi116和umc1044对(CML270×478)×CML270回交后代13份BCF6:7选系进行了抗性分子标记辅助选择。结果发现,2份选系与478具有1条相同带型,1份与478具有2条相同的带新型;另10份选系与CML270具有相同带型。人工接种鉴定结果表明,依据标记推断为不抗病的3份材料表现为感到高感纹枯病,而其余10份表现从感到高抗,带谱与病性指数相关系数达极显著水平,相关系数r=0.6625>r0.01=0.64,从中获得高抗玉米纹枯病选系478C4-2-2、478C4-2-3、478C4-2-4。配合力分析表明,478C4-2-2具有较高的一般配合力和特殊配合力,所配组合渝单24在2006年已通过重庆市审定。研究表明,标记鉴定和表型鉴定相结合,选择玉米纹枯病抗病自交系是完全有效的。     

茄子抗青枯病的遗传分析

茄子青枯病是由青枯雷尔氏菌侵染引起的一种主要土传病害,可使茄子植株萎蔫死亡、大量减产,严重制约着中国茄子的生产。为探明茄子抗青枯病的遗传规律,本研究以茄子高感病自交系“Rf”为母本,高抗病自交系“BC01”为父本,构建F₁、F₂、BC₁P₁、BC₁P₂世代遗传群体,采用伤根灌根接种法进行抗病性表型鉴定,利用植物数量性状主基因+多基因混合遗传模型分析方法对抗病遗传效应进行分析。结果表明,F₂代各病级株数呈现多峰、偏态分布;茄子青枯病的抗性遗传规律符合2MG-EAD模型,由2对主基因控制,感病为显性;主基因在F₂、BC₁P₁、BC₁P₂世代的遗传率分别为89.84%、43.19%、87.41%。研究结果为定位茄子抗青枯病主效基因,开发相关分子标记提供参考。     

玉米自交系及杂交种抗青枯病鉴定

1986-1989年采用自然鉴定和人工土壤接种鉴定的方法，对602份玉米材料的青枯病抗性进行了鉴定。人工接种鉴定397份，其中204份材料表现抗病。在正常年份，人工接种鉴定效果明显好于自然鉴定，但在重病区或青枯病流行年份，在尚未具备人工接种条件时，利用自然发病作为选育抗病材料仍有一定效果。玉米苗期接种青枯病重于成株期     

外来玉米种质的抗病和抗逆性选育

为改善国内玉米种质遗传基础狭窄、缺少优良抗性种质资源的状况,在500余份外引玉米材料中,鉴定筛选优良的抗病、抗逆基因,通过与国内骨干系杂交或组建抗病、抗逆群体,鉴别筛选含抗病、抗逆基因的材料回交纯合,再通过配合力测定,最终选育出新的优良自交系用于国内杂交种的选育,由此鉴定筛选出高抗锈病的材料25份、高抗大斑病10份、高抗穗腐病2份、高抗小斑病1份、高抗灰斑病2份、特早熟材料5份等,其中有现已标记定位出的不同抗大斑单基因的玉米材料8份,现已标记定位出的不同抗锈病基因的玉米材料25份,经进一步的遗传改良,获得超过35份抗病、抗逆自交系,利用新的抗逆自交系选育出特早熟玉米新品种植9831。     

番茄育种种质对南方根结线虫抗性评价及Mi-1基因检测

本研究以自育的26份番茄育种种质和抗南方根结线虫番茄品种‘仙客1号’及感南方根结线虫品种‘阿乃兹’为试验材料,采用病土盘栽自然发病鉴定、苗期二龄幼虫人工接种鉴定和PCR扩增及CAPS检测相结合的方法,研究供试材料对南方根结线虫的抗性水平及基因型。鉴定结果表明,供试的28份番茄材料中,高抗17份、抗病6份(包括抗病对照‘仙客1号’)、感病2份、高感3份(包括感病对照‘阿乃兹’),无免疫材料,且病土盘栽鉴定与苗期接种鉴定的结果一致。PCR扩增结果表明,27份番茄材料均能检测到一条750 bp的特异条带;TaqⅠ酶切结果显示,具有酶切位点的22份试材中有15份含纯合显性Mi-1/Mi-1基因、5份含杂合显性Mi-1/mi-1基因、2份含180 bp+250 bp+300 bp三条特异性片段,并且这22份试材对南方根结线虫表现高抗或抗,而其余5份不含显性Mi-1基因的试材则表现高感或感,证明Mi-1是具有抗南方根结线虫能力的基因,分子检测结果与病土盘栽鉴定和苗期接种鉴定的结果吻合率达92.6%;含有180 bp+250 bp+300 bp三条特异性片段的高抗材料可能含有其它抗根结线虫基因。分子检测与抗性鉴定相结合,能为创制抗南方根结线虫新种质提供一种更加省时、省力且经济有效的科学育种方法,可为番茄抗南方根结线虫育种提供理论依据和实践经验。     

长果种黄麻种质资源对黑点炭疽病抗性的鉴定

通过大田喷雾接种,对国内外77份长果种黄麻种质资源进行了黑点炭疽病(Collectotrichum gloeosporioides)抗性的鉴定,根据其病情指数确定反应类型,鉴定出高抗材料4份,中等抗病材料29份。     

野生花生抗青枯病种质的发掘及分子鉴定

以花生属5个区组的79份野生花生种质为材料,系统鉴定了野生花生对青枯病的抗性反应,从中发掘高抗青枯病的种质15份,含匍匐区组种质3份、直立区组1份、异形花区组1份、花生区组8份、未命名种质2份,抗病材料频率达到19%,高于栽培种花生资源的抗性频率。通过SSR分析表明,在所获得的抗青枯病野生花生材料中,四倍体野生种A.monticola与栽培种花生的亲缘关系最近,其次为花生区组的二倍体野生种A.duranensis和A.chacoense。根据DNA扩增结果,绘制了抗青枯病种质的指纹图谱,明确了其SSR分子特性。     

利用RIL群体创造抗黄曲霉兼抗青枯病的高油花生新种质

协同提高抗青枯病花生品种的黄曲霉抗性及含油量是我国花生育种的重要目标之一。利用远杂9102×中花5号杂交后代衍生的重组近交系群体(RIL),通过黄曲霉抗性、青枯病抗性鉴定及含油量测试,表明黄曲霉抗性受2对连锁并具累加作用主基因+加性多基因控制,含油量受2对具抑制作用主基因+加性多基因控制,RIL群体的黄曲霉抗性和含油量变异远远超过双亲的差异,表明它们均具有通过互补产生超亲性状的潜力。获得了抗黄曲霉或抗青枯病的高油后代家系18份,其中抗黄曲霉兼抗青枯病高油新种质1份(J091)。农艺性状和SSR分析结果表明,18份后代材料具丰富的遗传多样性,农艺性状优良,具有重要育种价值。     

Effects of resistance genes,heat tolerance genes and cytoplasms on expression of resistance to Pseudomonas solanacearum (E.F. Smith) in potato

Summary Effects of resistance genes and heat tolerance genes on expression of resistance to bacterial wilt caused by Pseudomonas solanacearum were investigated in 30 F₁ progenies from parents with different levels of bacterial wilt resistance and heat tolerance. A race 1 and a race 3 isolate of the bacterium were used for inoculation under screenhouse conditions at two locations. Results obtained indicated that with reduction in levels of parental resistance, resistance in the F₁ progenies was also reduced. Under hot conditions, a reduced heat tolerance in the genetic background also resulted in lower levels of resistance expression. The effect of heat tolerance tended to diminish at lower temperatures leaving the effect of resistance genes more consistent. There existed a strong interaction between resistance genes and genes for heat tolerance. The nature of resistance to bacterial wilt in potato and implications for breeding for resistance are discussed.Effects of reciprocal crosses on expression of resistance to a race 1 isolate under hot screenhouse conditions, were studied in 5 sets of reciprocal F₁ progenies involving different resistant and susceptible parents. The reciprocal differences observed were not significant suggesting absence of cytoplasmic effects on expression of resistance.     

Germplasm evaluation and transfer of Verticillium wilt resistance from Pima (<Emphasis Type="Italic">Gossypium barbadense</Emphasis>) to Upland cotton (<Emphasis Type="Italic">G. hirsutum</Emphasis>)

Verticillium wilt (VW, Verticillium dahliae) is a worldwide destructive soil-borne fungal disease and employment of VW resistant cultivars is the most economic and efficient method in sustainable cotton production. However, information concerning VW resistance in current commercial cotton cultivars and transfer of VW resistance from Pima (Gossypium barbadense) to Upland (Gossypium hirsutum) cotton is lacking. The objective of the current study was to report findings in evaluating commercial cotton cultivars and germplasm lines for VW resistance in field and greenhouse (GH) experiments conducted in 2003, 2006, and 2007. In the study, 267 cultivars and germplasm lines were screened in the GH, while 357 genotypes were screened in the field. The results indicated that (1) VW significantly reduced cotton yield, lint percentage, 50% span length and micronaire, but not 2.5% span length and fiber strength, when healthy and diseased plants in 23 cultivars were compared; (2) some commercial cotton cultivars developed by major cotton seed companies in the US displayed good VW resistance; (3) many Acala cotton cultivars released in the past also had good VW resistance, but not all Acala cotton germplasm are resistant; (4) Pima cotton possessed higher levels of VW resistance than Upland cotton, but the performance was reversed when the root system was wounded after inoculation; (5) VW resistance in some conventional cultivars was transferred into their transgenic version through backcrossing; and (6) some advanced backcross inbred lines developed from a cross between Upland and Pima cotton showed good VW resistance. The successful development of VW resistant transgenic cultivars and transfer of VW resistance from Pima to Upland cotton implies that VW resistance is associated with a few genes if not a major one.     

外源赤霉素浸种对茄子种子萌发和植株生长影响的研究

为研究赤霉素（GA3）对打破茄子种子休眠及植株生长的影响,以16份自交系和2份品种为试材,采用GA3处理种子,对不同自交系种子的休眠特性的解除和杂交品种的植株生长的影响进行研究。结果表明16份自交系取种后1个月芽率都较低,最高为25.5%,最低为0%,不同材料间存在极显著差异,说明初收茄子种子普遍存在休眠现象,但休眠深浅程度存在差异;经1000 mg/L GA3处理,所有自交系的芽率、芽势都有明显提高,其中6份芽率达到90%以上,说明GA3对解除茄子种子休眠效果极为显著。室温干燥器存储4个月后除06036和06039 2份亲缘来自中国北方的黑紫茄自交系芽率在低于80%,其他14份材料均在90%以上。利用GA3浸种处理‘农夫长茄’和‘庆丰紫红茄’对不同生长时期的植株、果实性状和产量调查发现,较高浓度GA3(2000 mg/L)对‘庆丰紫红茄’1叶1心期株高有极显著增加,低浓度GA3浸种处理对‘庆丰紫红茄’及不同浓度GA3对‘农夫长茄’的株高促进作用不明显;4叶期后,不同浓度的GA3浸种处理对‘农夫长茄’和‘庆丰紫红茄’植株高度、茎周径、叶片大小的影响差异均不显著,对2个品种的开花期、商品果性状（单果重、果长、果粗、畸形果率）及产量等影响均未达显著差异;且同一浓度浸种4 h与6 h对植株生长、商品果性状和产量的作用效果一致。本研究可为茄子商用种解除休眠提供理论依据。     

节瓜抗镰刀菌酸突变体的筛选和特性研究

枯萎病是节瓜的主要病害之一。为了创新节瓜抗枯萎病资源,利用枯萎病毒素镰刀菌酸（FA）为胁迫剂,以节瓜不定芽为筛选材料,进行了节瓜抗镰刀菌酸变异体离体筛选研究。试验结果表明,FA胁迫的适宜浓度为60~80mg&#8226;L-1。筛选出的抗性细胞系继代培养后仍保持对FA抗性,其再生植株的R1、R3代株系对节瓜枯萎病的抗性可以达到中抗至抗病级;与供体材料相比,再生系后代株系除枯萎病抗性增强外,其他主要农艺性状没有明显变化。表明利用抗FA细胞系离体筛选技术改良和创新节瓜抗枯萎病材料是可行的。     

转GHABF4转录因子棉花植株的耐盐性研究

AREB/ABFs转录因子家族基因主要参与干旱、高盐、低温等胁迫应答反应。为了获得具有较高耐盐水平的棉花新种质材料,通过农杆菌介导法将耐盐转录因子基因(GHABF4)导入陆地棉中棉35中,通过对转化植株的卡那霉素初步筛选及T1、T2、T3目的基因PCR的分子检测,获得T3转基因棉花纯合系。通过盐胁迫试验对5个T3转基因棉花株系和非转基因棉花对照进行耐盐性分析。结果表明,在200 mmol/L Na Cl胁迫下,与非转基因对照相比,5个转基因棉花株系株高提高2.5~4.4 cm,地上部分的鲜质量增加3.6%~11.8%;且抗氧化物酶SOD、POD、CAT活性以及叶绿素含量提高。在盐胁迫条件下,转GHABF4基因棉花表现出优良的生长和生理优势,转GHABF4基因能够提高棉花的抗盐能力。     

140份甜玉米自交系耐铝性鉴定

以华南农业大学选育的140份甜玉米自交系为材料,利用0.4 mmol·L-1的AlCl3溶液处理玉米幼苗,测定其相对根长、相对叶绿素含量、相对株高、相对鲜重和相对干重等值并汇总排序,进行耐铝性鉴定分析评价.结果表明,系统聚类分析将140份甜玉米自交系分为4个类群,类群Ⅲ中自交系平均耐铝水平较高.分析140个甜玉米自交系...     

The detection of QTLs controlling bacterial wilt resistance in tobacco (N. tabacum L.)

The bacterial tobacco wilt caused by Ralstonia solanacearum is one of the most destructive soil-borne diseases worldwide. One strategy to improve the resistance to bacterial wilt is to make use of plant varieties expressing wilt resistance genes. To characterize the genetics of wilt resistance and to identify relevant molecular markers for use in plant breeding, quantitative trait loci (QTLs) affecting tobacco bacterial wilt resistance were mapped in the F_2:3 and F_2:4 progeny produced from two crosses between the wilt-resistant breeding lines Enshu and Yanyan97 and the susceptible cultivar TI448A. A linkage map containing 118 loci in 24 linkage groups was constructed for 236 lines from the Enshu×TI448A cross, and a linkage map containing 96 loci in 24 linkage groups was constructed for 264 lines from the Yanyan97×TI448A cross. The wilt resistance of the progeny was examined in field trials conducted in Xuancheng, China, in 2010, 2011, and 2012. The disease severity was assessed on stems using separate rating scales. Mapmaker/EXP 3.0 and Mapmaker/QTL 1.1 were used to identify the qBWR-3a, qBWR-3b, qBWR-5a and qBWR-5b QTLs in linkage group 3 and 5; these four loci were strongly associated with resistance and explain 9.00, 19.70, 17.30, and 17.40 % of the variance in resistance, respectively. The close linkage of the markers PT20275 and PT30229 was detected in both the TI448A×Enshu and TI448A×Yanyan97 crosses, and this linkage group could be used to select individual resistant plants. These findings suggest that one strategy to combat bacterial wilt could be to exploit the resistance genes of the Enshu and Yanyan97 strains.     

转抗菌肽基因辣椒株系的青枯病抗性鉴定及系统选育

利用通过农杆菌介导技术获得的T0代转抗菌肽基因辣椒植株为试材,对其自交株系世代群体连续进行抗青枯病鉴定筛选、分子生物学检测和系统选育,首次获得了具有抗青枯病能力的转抗菌肽基因辣椒稳定株系。同时,对其主要果实性状、青枯病抗性进行的对比试验结果表明,转抗菌肽辣椒株系除抗青枯病能力明显提高外,果实性状基本不变。上述结果显示,外源目的基因主要特性的遗传是稳定的、表达是忠实的,从实践上验证了转抗菌肽基因工程操作的实用性。