中国马铃薯
中國馬鈴藷
중국마령서
CHINESE POTATO
2009年
5期
257-262
,共6页
陈勤%孙慧生%杨继良%李海燕%Sanjib Nandy%Debbie Beasley%王冬冬%Mark Goettel
陳勤%孫慧生%楊繼良%李海燕%Sanjib Nandy%Debbie Beasley%王鼕鼕%Mark Goettel
진근%손혜생%양계량%리해연%Sanjib Nandy%Debbie Beasley%왕동동%Mark Goettel
分子标记%抗晚疫病基因%墨西哥野生种%Solanum pinnatisectum
分子標記%抗晚疫病基因%墨西哥野生種%Solanum pinnatisectum
분자표기%항만역병기인%묵서가야생충%Solanum pinnatisectum
molecular mapping%late blight resistance gene%Mexican wild species%Solanum pinnatisectum
马铃薯晚疫病(Phytophthora infestans)和科罗拉多马铃薯甲虫(CPB)是马铃薯生产中最为严重的病虫害.培育高抗晚疫病和甲虫的马铃薯品种是加拿大马铃薯育种工作的重要组成部分.目前,我们实验室在二倍体IEBN墨西哥野生种中已鉴定出抗马铃薯晚疫病和甲虫的新基因,并利用原生质体融合技术成功的将其转移到栽培品种中.但是,培育出抗晚疫病和抗甲虫的马铃薯新品种仍然是一项艰难而繁杂的工作.为了加快分离抗性基因,建立与抗性基因紧密关联的DNA分子标记至关重要.本研究以感病的二倍体马铃薯品种S.cardiophyllum作为父本,与带有抗性基因的墨西哥野生种S.pinnatisectum杂交.用叶片离体鉴定的方法测试F1和BC1代群体的抗病性,从而筛选抗晚疫病和抗甲虫的植株.US-8/A2交配型病菌测试显示所有的F1代植株都表现出抗晚疫病,而在BC1群体中抗病与感病植株的比例为1:1.这个结果证明,在墨西哥野生种S. pinnatisectum中存在一个抗晚疫病的单显性基因Rpi1.马铃薯甲虫抗性检测中,BC1群体的抗虫性分离比例为1:3.这表明其对甲虫的抗性是由多基因遗传控制的.在F1和BC1群体中利用分子标记结合集团分离分析法(BSA)对S.pinnatisectum中的晚疫病抗性基因Rpi1进行精细作图.根据马铃薯第7条染色体上RFLP标记TG20A和CP56之间的EST和STS标记的序列信息,合成了27对特异性PCR引物.获得一些与抗晚疫病基因Rpi1相关联的新的DNA标记.对BC1群体中大量的个体植株进行的分析表明,在马铃薯第7条染色体上位于抗晚疫病基因Rpi1两侧的两个标记S1c9和GP127-300,它们与Rpi1基因的遗传距离分别为1.17 cM和3.89 cM.这些标记被用来筛选两个细菌人工染色体(BAC)文库,并分离出与晚疫病抗性相关的90-125 kb的BAC克隆,这些克隆将在后续的工作中通过图位克隆的方法而用于分离晚疫病抗性基因.同时分离与甲虫抗性密相关的分子标记的工作正在进行中.
馬鈴藷晚疫病(Phytophthora infestans)和科囉拉多馬鈴藷甲蟲(CPB)是馬鈴藷生產中最為嚴重的病蟲害.培育高抗晚疫病和甲蟲的馬鈴藷品種是加拿大馬鈴藷育種工作的重要組成部分.目前,我們實驗室在二倍體IEBN墨西哥野生種中已鑒定齣抗馬鈴藷晚疫病和甲蟲的新基因,併利用原生質體融閤技術成功的將其轉移到栽培品種中.但是,培育齣抗晚疫病和抗甲蟲的馬鈴藷新品種仍然是一項艱難而繁雜的工作.為瞭加快分離抗性基因,建立與抗性基因緊密關聯的DNA分子標記至關重要.本研究以感病的二倍體馬鈴藷品種S.cardiophyllum作為父本,與帶有抗性基因的墨西哥野生種S.pinnatisectum雜交.用葉片離體鑒定的方法測試F1和BC1代群體的抗病性,從而篩選抗晚疫病和抗甲蟲的植株.US-8/A2交配型病菌測試顯示所有的F1代植株都錶現齣抗晚疫病,而在BC1群體中抗病與感病植株的比例為1:1.這箇結果證明,在墨西哥野生種S. pinnatisectum中存在一箇抗晚疫病的單顯性基因Rpi1.馬鈴藷甲蟲抗性檢測中,BC1群體的抗蟲性分離比例為1:3.這錶明其對甲蟲的抗性是由多基因遺傳控製的.在F1和BC1群體中利用分子標記結閤集糰分離分析法(BSA)對S.pinnatisectum中的晚疫病抗性基因Rpi1進行精細作圖.根據馬鈴藷第7條染色體上RFLP標記TG20A和CP56之間的EST和STS標記的序列信息,閤成瞭27對特異性PCR引物.穫得一些與抗晚疫病基因Rpi1相關聯的新的DNA標記.對BC1群體中大量的箇體植株進行的分析錶明,在馬鈴藷第7條染色體上位于抗晚疫病基因Rpi1兩側的兩箇標記S1c9和GP127-300,它們與Rpi1基因的遺傳距離分彆為1.17 cM和3.89 cM.這些標記被用來篩選兩箇細菌人工染色體(BAC)文庫,併分離齣與晚疫病抗性相關的90-125 kb的BAC剋隆,這些剋隆將在後續的工作中通過圖位剋隆的方法而用于分離晚疫病抗性基因.同時分離與甲蟲抗性密相關的分子標記的工作正在進行中.
마령서만역병(Phytophthora infestans)화과라랍다마령서갑충(CPB)시마령서생산중최위엄중적병충해.배육고항만역병화갑충적마령서품충시가나대마령서육충공작적중요조성부분.목전,아문실험실재이배체IEBN묵서가야생충중이감정출항마령서만역병화갑충적신기인,병이용원생질체융합기술성공적장기전이도재배품충중.단시,배육출항만역병화항갑충적마령서신품충잉연시일항간난이번잡적공작.위료가쾌분리항성기인,건립여항성기인긴밀관련적DNA분자표기지관중요.본연구이감병적이배체마령서품충S.cardiophyllum작위부본,여대유항성기인적묵서가야생충S.pinnatisectum잡교.용협편리체감정적방법측시F1화BC1대군체적항병성,종이사선항만역병화항갑충적식주.US-8/A2교배형병균측시현시소유적F1대식주도표현출항만역병,이재BC1군체중항병여감병식주적비례위1:1.저개결과증명,재묵서가야생충S. pinnatisectum중존재일개항만역병적단현성기인Rpi1.마령서갑충항성검측중,BC1군체적항충성분리비례위1:3.저표명기대갑충적항성시유다기인유전공제적.재F1화BC1군체중이용분자표기결합집단분리분석법(BSA)대S.pinnatisectum중적만역병항성기인Rpi1진행정세작도.근거마령서제7조염색체상RFLP표기TG20A화CP56지간적EST화STS표기적서렬신식,합성료27대특이성PCR인물.획득일사여항만역병기인Rpi1상관련적신적DNA표기.대BC1군체중대량적개체식주진행적분석표명,재마령서제7조염색체상위우항만역병기인Rpi1량측적량개표기S1c9화GP127-300,타문여Rpi1기인적유전거리분별위1.17 cM화3.89 cM.저사표기피용래사선량개세균인공염색체(BAC)문고,병분리출여만역병항성상관적90-125 kb적BAC극륭,저사극륭장재후속적공작중통과도위극륭적방법이용우분리만역병항성기인.동시분리여갑충항성밀상관적분자표기적공작정재진행중.
Late blight (Phytophthora infestans) and Colorado potato beetle (CPB) are the most disastrous disease and insect problem of potatoes. Development of high levels of late blight and CPB resistant potatoes has become a high priority for the Canadian potato breeding program. New genes for late blight and CPB resistance have been identified in a wild 1EBN diploid Mexican species Solanum pinnatisectum which has been successfully transferred via protoplast fusion to the cultivated potato background. However, development of new potato cultivars with late blight and CPB resistance has been a difficult and cumbersome task. To accelerate the isolation of the resistant genes, molecular DNA markers tightly linked to the resistance are needed. In this study, a susceptible diploid potato S. Cardiophyllum was selected as the male parent to cross with S. Pin-natisectum. The F1 and BC1 populations were assessed for resistance to late blight and CPB by the detached leaf method. Dis-ease test using US-8/A2 mating type isolate revealed that all of the F1 individuals were resistant to the late blight. The ratio of late blight resistant plants to susceptible plants was 1 : 1 in BC1 populations. The results confirmed that a single dominant gene Rpil for late blight resistance was present in S. Pinnatisectum. For CPB resistance, a 1:3 resistant to susceptible ratio in BC1 populations confirmed polygenic inheritance. Molecular marker analysis combined with bulked segregating analysis (BSA) was carried out in the F1 and BC1 populations for fine mapping late blight resistant gene Rpi1 in S. Pinnatisectum. Twenty seven specific PCR primers were designed from sequence information of EST and STS markers located between RFLP markers TG20A and CP56 on potato chromosome Ⅶ. Several new DNA markers showed the linkage relation with the late blight resistant gene Rpi1. The analysis of a large number of individual plants from BC1 populations indicated that two mark-ers S1c9 and GP127～300 on chromosome Ⅶ are flanking the late blight resistant gene Rpi1 with a genetic distance of 1.17 cM and 3.89 cM, respectively. These markers were used to screen two bacterial artificial chromosome (BAC) libraries. Several BAG clones 90～125 kb in size linked with late blight resistance were isolated which will be used to isolate late blight resis-tance genes through the map-based cloning strategy. Development of molecular matkers closely linked with CPB resistance is on going.
