棉花黄萎病菌与抗黄萎病遗传育种研究进展

简要综述了棉花黄萎病菌及抗黄萎病遗传育种的研究进展。研究表明 ,各地的棉花黄萎病菌均存在致病力的分化 ,其致病机理是病菌侵入棉花后菌丝及孢子在导管内大量繁殖 ,同时刺激邻近的薄壁细胞产生胶状物质及侵填体而堵塞导管 ,使水分和养分运输发生困难 ,更重要的是病菌在棉株体内产生的糖蛋白毒素作用的结果。棉花抗黄萎病的遗传方式争论较大 ,但一般在温室由单一菌系接种鉴定时棉花黄萎病抗性表现为单基因遗传 ,而在田间病圃或用多菌系混合鉴定时 ,棉花黄萎病抗性表现为多基因遗传。由于陆地棉内缺乏高抗黄萎病资源 ,给棉花抗黄萎病育种带来一定困难 ,但 90年代以来 ,已育成 86 - 6、川 73 7、川 2 80 2、豫棉 1 9号、豫棉 2 1号等一些抗黄萎病的新品种。上述抗黄萎病品种在棉花黄萎病综合防治中起了重要作用     

Over-expression of Gastrodia anti-fungal protein enhances Verticillium wilt resistance in coloured cotton

Currently there are no adequate control measures for the cotton fungal diseases Verticillium wilt and Fusarium wilt, which are important factors limiting yield under certain conditions. In this study the gene encoding a Gastrodia anti‐fungal protein was introduced into three cultivars of coloured cotton using the method of pollen‐tube pathway transformation, with the purpose of obtaining transgenic plants with improved resistance to wilt. Of the 121 herbicide‐resistant cotton plants two, LB‐5‐8 and ZB‐1‐49, were scored as transgenic based on Southern blot, RT‐PCR analysis and in vitro anti‐fungal activity assay. Field analysis demonstrated that the transgenic lines LB‐5‐8 and ZB‐1‐49 possess an increased resistance to wilt. After 2 years of breeding, the progeny of LB‐5‐8 and ZB‐1‐49 lines still showed a stable and strong resistance to Verticillium wilt. Lines with high levels of resistance to Verticillium wilt obtained from the present study may be widely planted and help to reduce the future impact of cotton wilt on cotton production resulting in increased yields.     

接种不同致病力黄萎病菌的F1代棉花的基因型抗性研究

 在乌兹别克斯坦,棉黄萎病对棉花造成严重损失。为了选育能抗强致病力菌系的棉花品种,需确定棉花对不同地域的强致病力菌系的基因型抗性和F₁代的抗性遗传度。研究发现,品种Omad和品系L-44,L-408,L-155,L-1708对所选菌系的抗性最好。当受到侵染时,它们表现出超敏感性,但不表现黄萎病症状。F₁代的黄萎病抗性为超显性和显性遗传,且与鉴定方法无关。同时,存在中间性遗传。遗传优势度取决于其亲本的配合力、F₁代受侵染时的基因反应型和不同地域菌系的致病力。其中,Omad,С-5621,L-44,L-1708的表型抗性高;在F₁代中,组合L-155×С-5621和L1708×С-5621的表型抗性较高,在50％～80％。黄萎病抗性遗传控制的特点是趋向于最好或最差亲本的负或正的超显性、显性遗传以及中间型遗传。对所选菌系,杂交组合L-155×С-5621 和 L-155×L-44的综合抗性最好。     

Genotypic Resistance of F1 Cotton Hybrids by Inoculation with Different Virulent Isolates of the Fungus Verticillium Dahliae Klebahn

The plant pathogen Verticilliurn dahliae causes severe cotton losses in Uzbekistan.To create cotton varieties that are resistant to the more virulent races of V.dahliae we wanted to determine genotypic resistance of varieties and lines of cotton to more virulent isolates of different geographic populations of the fungus V.dahliae isolated from different cotton varieties in Uzbekistan and determine the inheritance of wilt resistance with hybrids of the first generation to various isolates of V.dahliae.     

Marker-assisted selection of Verticillium wilt resistance in progeny populations of upland cotton derived from mass selection-mass crossing

Two progeny populations of upland cotton derived from mass selection-mass crossing, M₃S₂F₅ and its family lines M₃S₂F_5:6, were generated from 17 hybrid cotton lines derived from regional trials conducted in the Yellow River basin and Yangtze River basin in China. These populations were used to verify 39 reported molecular markers that were related to quantitative trait loci (QTLs) for Verticillium wilt resistance of cotton. Only 12 of 39 markers were polymorphic; 19 had no polymorphisms, and amplification failed for eight markers. The differences in disease grades of aa/AA genotype individuals for five markers, BNL3241, NAU1225, NAU1230, JESPR153, and BNL3031, reached either significant or highly significant levels in at least one population. These markers can thus be effectively used for marker-assisted selection (MAS) of the target trait. Especially for JESPR153 and BNL3031, the differences in disease grades of aa/AA genotype individuals both reached either significant or highly significant levels in the two populations. These two markers should be given preferential consideration when undertaking MAS. The two flanking markers were more effective than the single flanking marker for MAS of single-loci QTL. The selection effect will be greatly enhanced through a reasonable allocation of marker combinations for multi-locus QTL polymerization. When using multi-locus markers for multi-locus QTL-assisted polymerization breeding, the selection effect can be improved progressively by increasing the number of polymerization markers. The possible interaction of different QTLs or genetic backgrounds does not influence the selection effect. A combination of resistant genotypes and disease grade performance enabled final selection of three individuals resistant to Verticillium wilt.     

棉花黄萎病菌菌体蛋白与其致病类型的关系

经液休培养获得５个棉花黄萎病菌不同致病类型的菌体培养物，并进行了菌体蛋白的十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析和氨基酸组成分分析。结果表明，美国落叶型Ｔ－９茵系的菌体蛋白电泳谱带数明显多于我国落叶型菌系ＶＤ－８菌系和３个不同致病类型的代表菌系，除安阳菌系外，其它４个菌系分别具有其蛋白特征带。我国落叶型ＶＤ－８菌系和强致病力类型泾阳菌系的菌体氨基酸含量明显高于中等致病力和弱致病力类型，落叶型和非落叶型菌系在某些氨基酸含量上具有明显差异。     

Inheritance of Dry Root Rot (Rhizoctonia bataticola) Resistance in Chickpea (Cicer arietinum)

The inheritance of resistance to dry root rot of chickpea caused by Rhizoctonia bataticola was studied. Parental F₁ and F₂ populations of two resistant and two susceptible parents, along with 49 F₁ progenies of one of the resistant × susceptible crosses were rested for their reaction to dry root rot using the blotting-paper technique. All F, plants of the resistant × susceptible crosses were resistant; the F₂ generation fitted a 3 resistant: 1 susceptible ratio indicating monogenic inheritance, with resistance dominant over susceptibility. F³ family segregation data confirmed the results. No segregation occurred among the progeny of resistant × resistant and susceptible × susceptible crosses.     

用相互嫁接和定量PCR分析棉花对棉花黄萎病的抗性

为了研究棉花对棉花黄萎病的抗性机制, 本文选用对棉花黄萎病表现抗病的海岛棉(Gossypium barbadense)材料海7124和Pima 90及感病的陆地棉(G. hirsutum)材料冀棉11, 通过相互嫁接的方法构建试验系统, 用棉花黄萎菌对其人工接种, 利用Real-time quantitative PCR (qPCR)技术分析其在感病和抗病棉花中侵染的差别。相互嫁接试验中感/抗和抗/感组合的病情指数介于感/感和抗/抗对照之间, 且相互嫁接棉株各个部位的IC值(侵染系数)也多介于其对照相应部位之间; 并且病情指数与不同部位IC值显著相关, 说明棉花黄萎菌可以通过接口在抗-感之间扩展。感/抗嫁接组合试验说明抗病材料的茎基部在抑制病原菌扩展中起重要的作用, 而抗/感类型试验说明抗病材料接口以上部分也具有抑制病原菌增殖的作用。总之, 抗病海岛棉无论作为砧木还是接穗, 都能有效抑制病原菌的扩展, 说明抗病海岛棉对棉花黄萎菌具全株抗性, 但茎基部在抑制病原菌扩展中起重要的作用; 同样, 感/抗和抗/感组合试验也说明感病材料的各个部位均不能抑制病原菌的定殖和扩展。     

陆地棉抗黄萎病性状的遗传及分子标记研究

 以陆地棉抗黄萎病品系常96和感病品种军棉1号为研究材料,构建了一个含有138个F₂单株的作图群体。利用强致病力菌株VD8对P₁、P₂、F₁、F_2：3群体进行营养钵接种,估算各世代的相对病指。应用主基因 多基因混合遗传模型分析得出该组合的最适遗传模型为C-0模型,即加性-显性-上位性多基因模型。对1998对SSR引物和230对SRAP引物进行筛选,获得148个SSR和6个SRAP多态性标记位点,进一步利用MAPMAKER作图软件,构建了一张含122个标记位点的陆陆杂种遗传连锁图。利用复合区间作图法在第9染色体NAU462-JESPR114区间内,检测到1个抗黄萎病QTL,可解释的表型变异为13.8%。     

棉花黄萎病抗性遗传和分子生物学研究进展

介绍了棉花黄萎病菌的致病机制 ,棉花黄萎病抗病机制及其遗传以及棉花黄萎病的鉴定方法 ,并对分子生物学在棉花黄萎病研究中的应用状况进展进行了综述。     

Genetic characterization of Gossypium arboreum accession PI 529740 for reniform nematode resistance

Breeding for reniform nematode (Rotylenchulus reniformis) resistance is hindered by the lack of resistance in upland cotton (Gossypium hirsutum) cultivars. Resistance has been frequently identified in accessions from the Gossypium arboreum germplasm collection with accession PI 529740 rated as highly resistant. Accession PI 529740 was crossed with the susceptible G. arboreum accession PI 529729 to develop an F₂ population for genetic characterization. The population showed quantitative variation suggesting multiple genes conferred the resistant phenotype. Thirteen of the 216 F₂ plants showed resistance similar to the resistant parent and these data supported a two recessive gene model. Sixty plants were classified as resistant or moderately resistant, indicating a single recessive gene conferred the moderately resistant phenotype. The classification of 24 F_2:3 families for nematode resistance generally supported the classification of the corresponding F₂ plants; however, most families were highly variable for infection with no families rated as resistant. This information will aid in the introgression of resistance into upland cotton as larger populations will be required to successfully recover resistance conferred by multiple recessive genes.     

Analysis of mode of inheritance of Fusarium wilt resistance in castor (Ricinus communis L.)

Castor (Ricinus communis L.) is an important industrial oilseed crop grown worldwide. Wilt caused by Fusarium oxysporum f.sp. ricini is a devastating disease in castor. The inheritance mode of wilt resistance was investigated. The F₁, F₂ and backcross generations of four crosses involving four resistant and three susceptible parents were developed. The role of digenic (R¹ and R²) epistatic interactions on wilt resistance was confirmed. The 15 : 1, 9 : 7 and 13 : 3 ratios indicated duplicate dominant, duplicate recessive and dominant and recessive epistatic interactions, respectively. Castor parents used in the crosses exhibited varied inheritance modes. All generations of a cross exhibited similar inheritance mode when parents were comparable. However, generations varied in inheritance mode when parents were not comparable in inheritance mode. These results would have practical interest when decisions are required regarding the choice of parents and methodology in resistance and hybrid breeding. The results also provided a basis for investigating molecular genetics of wilt resistance mechanisms.     

棉花种间杂交渐渗系抗黄萎病性状遗传分析

利用抗黄萎病棉花种间渐渗系冀79作父本,高感病陆地棉品系1096作母本,配制杂交组合。感病对照、亲本及F1、F2在田间混生病圃鉴定。对亲本、F1及F2分离群体的抗性表现进行调查、统计,分析该抗源抗黄萎病性状的遗传方式,结果表明,该抗源抗性是由1个显性抗(耐)病基因和2个加性基因共同起作用,其中加性基因起主要作用。并且2个加性基因是独立遗传的,当2个加性基因同时存在并且纯合时,植株表现为高抗;当2个加性基因同时存在并且杂合时,植株表现为抗病;当只有1个加性基因存在时,不论纯合还是杂合,植株都表现为耐病;当2个加性基因不存在时,植株表现为感病。     

棉花黄萎病菌致病类型研究

以研究手段为主线，从形态学特征、凝胶电泳、血清学反应、营养体亲和性及几项分子生物学技术等方面讨论了目前国内外在划分棉花黄萎病菌种内致病类型上的研究进展状况，并在本文基础上作了总结。     

新疆南部棉区黄萎病菌种群致病性分化及变异

 采用鉴别寄主法和特异性引物PCR检测技术对采自新疆南部（南疆）棉区的36个棉花黄萎病菌菌系进行检测,以期明确其病原种群致病性分化及变异情况。鉴别寄主法测定结果显示：供试菌系中致病性强、中、弱的菌系分别为27个、4个和5个,各占75.0%,11.1%和13.9%。特异性引物（ND1/ND2和D1/D2）PCR检测结果显示,供试36个菌系中落叶型菌系12个,占33.3%;非落叶型菌系24个,占66.7%。经统计,落叶型菌系的致病力明显高于非落叶型菌系,其平均病指分别为43.8和31.8。接种试验证明,无论落叶型菌系和非落叶型菌系,接种后都可产生落叶症状,但落叶的情况有明显差别,落叶症状的轻重不仅与菌系的致病类型有关,还与棉花品种的抗病性密切相关。     

小麦品系天95HF2抗叶锈基因定位

苗期基因推导表明,小麦品系天95HF2高抗我国目前多数叶锈菌生理小种。为了确定这一品系所携带的抗病基因,以天95HF2和感病小麦品种郑州5389杂交,获得F1和F2代群体,用叶锈菌小种FHTT和PHTS分别对双亲及其杂交后代进行叶锈抗性鉴定并进行分子标记分析。结果表明,用叶锈菌小种FHTT接种F2代群体时呈现1对显性基因的抗感分离比例,经过亲本和抗感池间标记筛选以及F2代群体的标记检测,Lr1的STS标记WR003和位于5DL的SSR标记wmc443与该抗病基因连锁,遗传距离分别为2.9cM和3.1cM,根据抗性特点和染色体位置推断该基因可能为Lr1。用叶锈菌小种PHTS接种F2代群体时呈现2对基因的抗感分离,分子标记分析结果表明,其中一个基因为Lr1,另一个基因可能为LrZH84。     

陆地棉植株组织结构和生化代谢与黄萎病抗性的关系

通过对8个陆地棉品种的植株组织结构、酶活性和根系分泌物的比较分析,研究陆地棉黄萎病抗性机制。研究结果表明,陆地棉抗病品种根和茎的导管细胞壁厚、直径小、数目多,髓射线数目多、单位面积薄壁细胞数多,有助于抵御棉花黄萎病菌的侵入与扩展。过氧化物酶（POD）活性与棉花抗黄萎病的关系不明显,但苯丙氨酸解氨酶（PAL     

Genetic analysis of resistance to leaf-rust (Puccinia recondita f. sp. tritici) pathotypes in the durum wheats 'PBW 34' and 'DWL 5023'

Genetic analysis of leaf-rust resistance was conducted on two durum wheats. Triticum durum cvs. ‘PBW 34’ and ‘DWL 5023’ were crossed with the leaf-rust-susceptible durum wheat ‘Malvi Local’. The F₁, F₂ and F₃ generations were tested against leaf-rust pathotypes 1, 77A and 108. In ‘PBW 34’, a single dominant gene was effective against each of the pathotypes 1 and 108, whereas two independently inherited dominant genes were effective against pathotype 77A. In ‘DWL 5023’, two independently inherited dominant genes were operative against pathotypes 1 and 77A, whereas a single dominant gene was identified as being operative against pathotype 108. Allelic tests on F₂ generation and joint segregation analysis on F₃ generation seedlings, suggested that two different genes in each cultivar are effective against these three leaf-rust pathotypes. Cultivar ‘PBW 34’ has Lrd1 and Lrd2 genes whereas Lrd1 and Lrd3 genes are present in ‘DWL 5023’.     

棉花黄萎病菌与品种互作的格局分析

 以23个棉花品种(系)接种18个黄萎病菌株的抗感表现(病情指数)为分析对象,在聚类分析的基础上用双标图方法研究了寄主 病原的互作格局。结果表明：23个棉花品种分为6个抗病性不同的品种组,18个菌株聚为5个致病力不同的菌株群。5031和50312组成的品种组G1对所有菌株群具有最高抗性,可作为抗黄育种的高抗质源。来自河北成安的VD25与来自江苏常熟的XS4、XS6和XS7组成的菌株群S4与G5品种组存在互作,其致病力最强,是棉花黄萎病的优势菌株群,在抗黄育种和抗黄鉴定中应予以高度重视。S2和S3菌株群与56%的参试品种构成的3个品种组间存在互作,是潜在的优势菌株群。具平均抗病性的G2品种组是黄萎病菌的优良寄主。     

Inheritance of resistance to angular leaf spot in common bean and validation of the utility of resistance linked markers for marker assisted selection out side the mapping population

Inheritance of resistance to angular leaf spot (ALS) disease caused by Phaeoisariopsis griseola (Sacc.) Ferr was investigated in two common bean cultivars, Mexico 54 and BAT 332. Both Andean and Mesoamerican backgrounds were used to determine the stability of the resistance gene in each of the two cultivars. Resistance to P. griseola was phenotypically evaluated by artificial inoculation with one of the most widely distributed pathotypes, 63–39. Evaluation of the parental genotypes, F₁, F₂ and backcross populations revealed that the resistance to angular leaf spot in the cultivars Mexico 54 and BAT 332 to pathotype 63–39 is controlled by a single dominant gene, when both the Andean and Mesoamerican backgrounds were used. Allelism test showed that ALS resistance in Mexico 54 and BAT 332 to pathotype 63–39 was conditioned by the same resistance locus. Resistant and susceptible segregating populations generated using Mexico 54 resistant parent were selected for DNA extraction and amplification to check for the presence /absence of the SCAR OPN02 and RAPD OPE04 markers linked to the Phg-2 resistance gene. The results indicated that the SCAR OPN02 was not polymorphic in the study populations and therefore of limited application in selecting resistant genotypes in such populations. On the other hand, the RAPD OPE04 marker was observed in all resistant individuals and was absent in those scored susceptible based on virulence data. Use of the RAPD OPE04 marker in marker-assisted selection is underway.