甘蓝型油菜显性细胞核雄性不育基因的AFLP标记

用甘蓝型油菜双基因显性细胞核雄性不育系Rs1046A和欧洲油菜品种Samourai构建了一个回交分离群体。在群分法（BSA）构建的不育池和可育池中共筛选了256对AFLP引物组合,找到了与不育基因紧密连锁的两个AFLP标记(EA03MC1599和EA07MC01235), 它们与不育基因的遗传图距分别是3.5 cM和5.5 cM,而且位于不育基因的同一侧,标记间相     

利用分子标记辅助选育白菜薹复等位基因型雄性不育系

为解决白菜薹杂种优势利用中的杂交制种手段问题,以大白菜复等位基因型雄性不育系为不育源,设计定向转育方案,采用连续回交的方法转育不育性和农艺性状,同时利用分子标记辅助选择目标基因型植株,向白菜薹自交系中转育不育基因,创制白菜薹雄性不育系。通过26对SSR引物的筛选鉴定,获得与显性雄性不育基因Ms紧密连锁、且同样可以标记同一位点恢复基因Msf和可育基因ms的分子标记GSSR1。经过3个世代的回交转育,创制出了具有100%不育度和100%不育株率的白菜薹复等位基因型雄性不育系BGMS-3。以BGMS-3为母本,与6个白菜薹自交系杂交,筛选出1个强优势组合C3。     

辣椒CMS恢复基因的遗传分析及基因定位

本研究以辣椒胞质雄性不育系131BC5A与恢复系139D-21-3为亲本,构建了包含210个单株的F2群体,采用CAPS、SSR及SCAR等分子标记技术,对辣椒胞质雄性不育恢复基因进行遗传分析和基因定位研究。田间调查和遗传分析结果表明,F2群体的表现型中可育与不育的分离比为3:1。采用分离群体分组分析法(BSA),从F2可育群体和不育群体中各随机选取10株,构建可育和不育基因池。研究选用295对引物在亲本间进行多态性筛选,其中43对引物在亲本间表现出多态。通过进一步分析43对引物在基因池间的多态性,筛选出Rf基因连锁标记14个。然后对F2群体的210个单株进行连锁分析,最后将恢复基因定位在SSR标记pep43和pep20之间,约3.9 c M(或498.6 kb)的区间内,与两个标记的遗传距离分别为1.3 c M与2.6 c M。本研究为Rf基因的精细定位和克隆奠定了良好基础,也为辣椒雄性不育恢复系的分子标记辅助选择育种提供了理论参考。     

利用红麻HcPDIL5-2a非全长基因创制雄性不育新种质

为创造转基因雄性不育种质,将缺失酶活性中心序列的红麻HcPDIL5-2a非全长基因通过花粉管通道法转化红麻保持系722B,获得了海南冬繁不育、南宁夏繁可育的低温敏雄性不育种质722THS,并从其姊妹交后代选育出了稳定型核不育(GMS)系722HS;此后,又以722HS作母本,以非转基因野生型722B为轮回亲本连续回交,选育出了细胞质雄性不育系722HA。对722THS与722HA的细胞学观察和线粒体DNA分子鉴定表明,二者的小孢子败育时期均为双核期,但722HA发生了线粒体DNA重排,而722THS与722B的线粒体DNA保持不变。本研究结果为转基因创造作物雄性不育种质找到了一条新途径,具有重要的科学意义和广阔的应用前景。     

与甜椒隐性细胞核雄性不育基因紧密连锁的SRAP分子标记的研究

以甜椒隐性细胞核雄性不育两用系AB91为材料,采用混合集群分析法(BSA)构建了不育池与可育池.利用SRAP分子标记技术,筛选了225对SRAP引物组合及1393对EcoR Ⅰ和Mse Ⅰ引物组合,获得了与隐性核不育基因连锁的2个SRAP标记:E37 M39、E44M93,片段长度约为200 bp和500 bp,与育性...     

谷子显性雄性不育基因Msch的AFLP标记

利用雄性不育是实现谷子杂种优势利用最经济、有效的途径之一.为了寻找与不育基因Msch紧密连锁的分子标记,提高不育系的选育效率,本研究构建了Msch不育/可育近等基因系(NILs),通过对400对AFLP引物组合进行筛选,找到了与不育基因紧密连锁的两个AFLP标记(P17/M37224和P35/M52208),与不育基因的遗传距离分别是2.1 cM和1.4 cM,而且位于不育基因的同一侧,标记间相距0.7 cM.这两个AFLP标记可有效用于分子标记辅助选择育种.     

甘蓝型油菜细胞质雄性不育恢复基因的SSR标记及初步定位

以甘蓝型油菜恢复系Q 16C和不育系2116A为亲本构建F2群体,根据BSA法的构建原则,使用F2群体分别构建了可育基因池与不育基因池。运用98对SSR引物对油菜亲本及基因池进行了多态性分析,结果表明,有1对引物(OS 31)能在亲本和基因池间表现出多态性,使用该引物继续对F2群体的230个单株进行分析,表明标记(OS 31)与恢复基因Rfp的遗传距离为0.08cM,可作为恢复系辅助育种的候选标记。     

菜薹雄性不育相关基因的ISSR分子标记筛选

菜薹(Brassica campestris L.ssp.Chinensis var.utilis Tsenet Lee)属于十字花科芸薹属,原产中国,是华南地区重要的特产蔬菜之一.利用分子标记技术进行菜薹雄性不育相关基因的定位对菜蕞品质创新和选育新品种具有重要意义.目前尚未见有利用ISSR分子标记技术进行菜薹雄性不育分析的研究报道.利用ISSR技术对菜薹雄性不育系及其保持系的基因组DNA进行比较分析:结果显示100个ISSR引物中,只有引物UBC843的扩增结果在两系之间表现出了差异性,找到了不育系和保持系间的特异扩增片段.回收该特异扩增片段并进行克隆测序,测序结果表明该片段全长为462 bp.与白菜其它育性相关序列比较,同源性均低于50%.根据测序结果设计了一对特异引物,将其转化为更稳定的SCAR标记.经F2群体验证,ISSR引物扩增得到的特异片段很有可能来自核DNA.     

一个辣椒胞质雄性不育SCAR标记的KASP转化及其应用

为了在辣椒三系杂交育种研究中缩小群体筛选范围,减小工作量,提高不育系和保持系选择效率,根据细胞质育性标记SCAR130的序列多态性位点设计KASP标记引物,使其转化为KASP130分子标记,并以辣椒三系材料的雄性不育系、保持系、恢复系和F_1杂交种为试验对象,利用荧光定量PCR仪LC480和LGC公司的SNPline这两种检测平台将KASP130标记应用于辣椒细胞质类型检测,并对该标记稳定性和可靠性进行了检验。结果表明, KASP130标记同SCAR130标记一样可以把待试辣椒材料准确地分为可育细胞质(N)和不育细胞质(S),并在分子标记辅助选择育种中成功应用到辣椒细胞质育性的早期鉴定以及辣椒保持系和雄性不育系的回交育种研究。综上,细胞质育性标记SCAR130已经被成功转化为KASP130分子标记,该标记可以明确鉴定辣椒的细胞质类型,这为其在辣椒三系杂交育种上的应用奠定了研究基础。     

奶白菜核基因雄性不育系的选育与利用

为解决奶白菜杂交种生产中的杂交制种手段问题,配制优良杂交种.以核不育“复等位基因遗传”假说为指导,以青梗白菜核基因雄性不育系00S107作不育源,采用连续回交转育同时测交鉴定基因型的方法,定向转育奶白菜核基因雄性不育系.并利用转育成的不育系与奶白菜优良自交系配制杂交组合,进行了杂种优势分析.选育出园艺学性状与目标品系相似,具有100％不育株率和不育度的奶白菜核基因雄性不育系GMS3,筛选出2个产量高、整齐度高的优异杂交组合GMS3×B1、GMS3×B2.定向转育模式兼顾了不育基因和园艺学性状的转育,解决了奶白菜核不育系转育和利用的难题.     

白菜型油菜(B. campestris L.)双显性核不育896AB的选育

以北方白菜型油菜自交系Ｓ３－１８为母本,加拿大白菜型春油菜Ｔｏｂｉｎ为父本杂交,在自交２代中出现不育株３７ＡＢ,经研究认为,该不育材料是由２种不育基因（ＭＳＭＳｒｆｒｆ,ＭＳｍｓｒｆｒｆ）,４种保持基因（ＭＳＭＳＲｆｒｆ,ＭＳｍｒＲｆｒｆ,ｍｓｍｓＲｆｒｆ,ｍｓｍｓｒｆｒｆ）组成的杂合不育群体,用成对保持,双隐性测交的方法,经２～３个世代的选育,育成８９６ＡＢ双显性不育系及１２５ａｂ等双隐性临保系     

Identification of a SCAR marker linked to a recessive male sterile gene (Tems) and its application in breeding of marigold (Tagetes erecta)

In marigold, an F₂ segregation population of 167 plants was constructed from a cross of a line (M525A) carrying the male sterility trait × an inbred line (f53f). In line M525A, the male sterility trait was controlled by the recessive gene, Tems . The intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) techniques combined with bulked segregant analysis were used to develop markers linked to the trait. From a survey of the 38 ISSR primers and 170 SRAP primer combinations, only one SRAP marker that was closely linked to the target trait was identified and successfully converted into sequence characterized amplified region (SCAR) marker that was located within 2.4 cM from Tems locus. The marker was validated with five other two-type lines and in each case the male fertile plants were reliably identified. This SCAR marker therefore permits the efficient marker-assisted selection of male sterile individuals in breeding programmes of marigold and will greatly facilitate the breeding of F₁ cultivars.     

Modified complementation test of male sterility mutants in pepper (Capsicum annuum L.): preliminary study to convert male sterility system from GMS to CMS

The male sterility system in hybrid seed production can eliminate the cost of emasculation and ensure seed hybridity through avoidance of self pollination. GMS and CMS are two types of male sterility system that currently employed in pepper breeding. Conversion from GMS to CMS will increase the male sterility proportion of female parent from 50 to 100%. In this study, segregation analysis of four male sterile mutants consisting of one CMS mutant (CA1) and three GMS mutants (GA1, GA3 and GA4) showed that each had single recessive gene inheritance. A modified complementation test was performed by replacing male sterile mutants with their maintainer line as male parent. The nuclear restorer gene for CMS was independent of all nuclear restorer genes for GMS and all nuclear restorer genes for GMS were independent each other. Further observation on CMS and GMS male sterility loci revealed that GA1 and GA3 had mutated in both nuclear restorer genes for CMS and GMS, while CA1 and GA4 each carried mutation in single male sterility system of nuclear restorer gene for CMS and GMS, respectively. Conversion from GMS to CMS in the case of lines carried mutations in both sterility systems required only S-type cytoplasm donor, while lines carried mutation in single nuclear restorer gene for GMS required not only S-type cytoplasm but also rf allele donors. The important finding is the broader function of maintainer line in certain male sterility system that can be used as a maintainer or restorer line for other male sterility systems. We also confirmed that line CC1 is the general restorer for both CMS and GMS systems.     

利用分子标记辅助选育大白菜核基因雄性不育系

本研究以含不育基因Ms的大白菜细胞核复等位基因型雄性不育两用系"AB01"的可育株(MsfMs)为供体亲本,以高代自交系‘a20’(msms)为轮回亲本,采用杂交和连续回交转育方法,利用与不育基因Ms连锁的SCAR标记syau_scr01辅助不育基因Ms选择,成功地将不育基因转育到可育品系‘a20’中,育成了不育度和不育株率均为100%,植物学性状与自交系‘a20’相近的新核不育系GMS4。选择结果表明syau_scr01选择的准确率为100%,验证了该标记可以用于大白菜核不育系转育辅助选择。     

Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus)

Rs1046AB is a genic male sterile two‐type line in rapeseed that has great potential for hybrid seed production. The sterility of this line is conditioned by the interaction of two genes, i.e. the dominant genic male sterility gene (Ms) and the suppressor gene (Rf). The present study was undertaken to identify DNA markers for the Ms locus in a BC₁ population developed from a cross between a male‐sterile plant in Rs1046AB and the fertile canola‐type cultivar ‘Samourai’. Bulked segregant analysis was performed using the amplified fragment length polymorphism (AFLP) methodology. From the survey of 480 AFLP primer combinations, five AFLP markers (P10M13₃₅₀, P13M8₄₀₀, P6M6₄₁₀, E7M1₂₃₀ and E3M15₁₀₀) tightly linked to the target gene were identified. Two of them, E3M15₁₀₀ and P6M6₄₁₀, located the closest, at either side of Ms at a distance of 3.7 and 5.9 cM, respectively. The Ms locus was subsequently mapped on linkage group LG10 in the map developed in this laboratory, adding two additional markers weakly linked to it. This suite of markers will be valuable in designing a marker‐assisted genic male sterility three‐line breeding programme.     

广亲和粳稻恢复系选育方法

广亲和粳稻恢复系的选育除采用ＷＡ型恢复与广亲和粳稻杂交选育和不同广亲和恢复系间杂交选育外,本研究提出以粳稻为父本杂交选育的４种办法：１、以型杂交稻Ｆ１为母本转育同质广亲和恢复系;２、以ＷＡ型下育系为母本筛选同质广亲和恢复系;３、以ＷＡ型广亲和不育系为母本,筛选同质广亲和恢复系;４、以同质广亲和恢复系为母本转育新的恢复系。用上述方法已育成０２４２８等广亲和恢复系,并可同时恢复ＷＡ型和ＢＴ型胞质不育系     

质体转化开创作物杂种优势利用新途径

普通核雄性不育性能够满足对理想不育系选育的要求，是水稻等作物杂种优势利用的理想遗传元件。只要能解决其不育系繁殖问题，是理想的杂种优势利用方式。通过普通核雄性不育性的可育基因质体转化，将可育基因转移到存在于细胞质中的质体基因组中，可创造普通核雄性不育系的保持系，繁殖出普通核雄性不育系，从而实现三系法利用杂种优势；利用可产生雄性不育性的基因如TA29-barnase通过质体转化，向质体基因组中转移，产生由细胞质中转基因雄性不育基因控制的雄性不育系，任何常规品种都能作为其保持系，转barstar可育基因系(或常规品种)作为恢复系(或父本)，可同样实现三系法利用杂种优势。上述两种途径都是创造植物杂种优势利用的新途径。     

桔黄大牛角椒的果皮色遗传及雄性不育系的转育研究

为选育黄果皮牛角新品种,利用本课题组转育成的牛角椒红果皮雄性不育系、红果皮保持系、美国大牛角高代自交系探讨生理成熟时牛角椒果皮颜色的遗传规律、牛角椒雄性不育性与果皮颜色的遗传规律。结果表明：牛角椒的果皮颜色的遗传,由1对等位基因控制,红色对黄色为显性遗传;牛角椒雄性不育性与果皮颜色的遗传属独立遗传。以此为依据提出了以黄果皮保持系材料为轮回亲本,转育黄果皮牛角椒雄性不育系的遗传模式,为转育黄果皮牛角椒新品种打下理论基础。     

Directional transfer of a multiple-allele male sterile line in Brassica campestris L. ssp. chinensis (L.) Makino var. rosularis Tsen et Lee

To produce hybrid seeds of Wutacai (Brassica campestris L. ssp. chinensis (L.) Makino var. rosularis Tsen et Lee), a “directional transfer program” was designed to breed the multiple-allele male sterile line of Wutacai. A multiple-allele male sterile line of Naibaicai (Brassica campestris L. ssp. chinensis L., S01) was used as the male sterile resource, and an inbred line of Wutacai (WT01) was used as the target line. Recurrent backcrossing was employed to transfer the male sterility and other botanical traits simultaneously, while the genotype was identified through test crossing. The male sterility was successfully transferred from S01 to WT01. A new male sterile line, GMS-3, with similar botanical traits to WT01, was bred. Four hybrid combinations were generated with GMS-3 as the female parent. One hybrid (C1) that contained the most desirable traits was developed from the new male sterile line.