运用分子标记辅助选择技术改良9311白叶枯病抗性的研究

以9311为轮回亲本，以IRBB21为抗白叶枯病供体亲本，通过杂交和三次回交以及两次自交，结合农艺性状的选择，建立起具44个株系的BeF3群体。分别用连锁标记pTA248和基因内STS标记MXA21对抗性基因Xa21进行前景选择。用均匀分布在水稻全基因组的248个SSR标记，进行两亲本之间的多态性分析，获得76个具多态性的SSR标记，用于背景选择。结果选到基因型背景回复到轮回亲本的73．68％-88．16％，且Xa2j基因纯合的株系4个，分别为M071、M082、M086和M087。以Xa21的鉴别菌系菲律宾6号小种PxO99接种鉴定表明，9311为感(S)，IRBB21为抗(R)，中选株系M071和M082的抗性与IRBB21相同，为抗(R)级，M086和M087抗性略低于IRBB21，为抗到中抗(R／MR)，这两个株系内单株之间抗性有差异。农艺性状考察结果显示，中选株系与轮回亲本9311相似。以中选株系M086与培矮64S配制的F1和原组合两优培九(培矮64S/9311)进行比较，对白叶枯病的抗性有了明显提高，综合农艺性状相似。论文就白叶枯病抗性基因的综合利用以及育种方法进行了讨论。     

运用分子标记辅助选择技术改良9311白叶枯病抗性的研究

以9311为轮回亲本,以IRBB21为抗白叶枯病供体亲本,通过杂交和三次回交以及两次自交,结合农艺性状的选择,建立起具44个株系的BC3F3群体.分别用连锁标记pTA248和基因内STS标记MXA21对抗性基因Xa21进行前景选择.用均匀分布在水稻全基因组的248个SSR标记,进行两亲本之间的多态性分析,获得76个具多态性的SSR标记,用于背景选择.结果选到基因型背景回复到轮回亲本的73.68%-88.16%,且Xa21基因纯合的株系4个,分别为M071、M082、M086和M087.以Xa21的鉴别菌系菲律宾6号小种PXO99接种鉴定表明,9311为感(S),IRBB21为抗(R),中选株系M071和M082的抗性与IRBB21相同,为抗(R)级,M086和M087抗性略低于IRBB21,为抗到中抗(R/MR),这两个株系内单株之间抗性有差异.农艺性状考察结果显示,中选株系与轮回亲本9311相似.以中选株系M086与培矮64S配制的F1和原组合两优培九(培矮64S/9311)进行比较,对白叶枯病的抗性有了明显提高,综合农艺性状相似.论文就白叶枯病抗性基因的综合利用以及育种方法进行了讨论.     

云南高原粳稻抗白叶枯病新品系云资抗21号的选育

以高产优质的云南高原粳稻品种滇系4号为轮回亲本，以IRBB21为白叶枯病抗性基因Xa21的供体亲本，通过杂交和3次回交以及5次自交形成BC3F，代材料，并结合后代材料的白叶枯病抗性鉴定及农艺性状的评价选择，选育出高原粳稻新品系云资抗21号，对云资抗21号的21个单株采用与Xa21连锁标记RM21和RM229、以及22个独立分离的SSR标记进行检测，结果表明这21个单株在RM21位点上均为IRBB21的纯合基因型，在RM229位点上有1个单株显示为轮回亲本的纯合基因型，其余的20个单株均为IRBB21的纯合基因型，而在其余22个SSR标记位点上均显示为轮回亲本的纯合基因型。该新品系白叶枯病抗性与IRBB21相近，农艺性状与轮回亲本相似，具有很好的应用前景。     

分子标记辅助选育抗稻白叶枯病的低温敏核不育系3178S

本研究以IRBB21为Xa21基因供体，籼型低温敏不育系399S为低温核不育系基因供体，采用杂交和回交方法，逐代用分子标记检测手段，导入广谱抗白叶枯病基因Xa21和低温敏核不育系基因，聚合双亲的有利性状，育成抗白叶枯病的籼型低温敏核不育系3178S，其抗性达到了IRBB21的抗性水平，且保持了399S稳定的育性和双亲的优良经济性状。     

分子标记辅助选择Xa23基因培育杂交稻抗白叶枯病恢复系

水稻抗白叶枯病新基因Xa23抗谱广、抗性强,被定位在第11染色体上。以携带抗白叶枯病基因Xa23的抗病品系CBB23为抗源,以优良杂交稻亲本9311和1826为受体材料,采用杂交和复交,在分离群体中利用与Xa23紧密连锁的EST标记C189进行分子标记辅助选择,通过苗期分子标记检测和成株期农艺性状选择,获得160份目标基因纯合且农艺性状稳定的株系。使用鉴别菌系P6,采用人工剪叶接种法,在苗期和孕穗期对21份重点株系进行了抗性鉴定,所有株系在苗期和孕穗期都表现抗病。同时对3个不育系与其中5个株系配制的15个杂交组合进行了抗性鉴定,所有组合在苗期表现抗或中抗,在孕穗期表现抗。对其中6个杂交组合进行了品比试验,2个组合产量略低于对照两优培九,其余4个组合的产量均高于两优培九,用于测配的3个株系C6201、C6271和C6351有望作为杂交稻恢复系在生产中应用。研究表明在育种进程中利用与Xa23基因紧密连锁的分子标记C189开展抗白叶枯病分子标记辅助育种是一种有效的途径。     

应用分子标记辅助选择培育抗稻白叶枯病光敏核不育系3418S

分子标记辅助选择(MAS)因其对目标基因的快速准确选择为回交育种提供有效工具.本研究以回交转育为基础,逐代用分子标记检测导入广谱抗白叶枯病基因Xa21,在保持光敏核不育系3418S原有优良性状和稳定育性的基础上,增强其抗白叶枯病的能力.育成抗白叶枯病的籼型光敏核不育系3418S.其抗性达到了IRBB21的抗性水平,且保持了3418S的     

分子标记辅助选择Xa23基因改良杂交稻亲本的白叶枯病抗性

白叶枯病是世界上影响水稻产量最严重的病害之一,本研究利用携有目前已知的、抗谱最广和抗性最强的显性基因Xa23的水稻材料CBB23作为基因供体,以感白叶枯病的杂交水稻的3个骨干亲本培矮64S、明恢86和C418作为受体,采用杂交和复交,在分离群体中利用与Xa23紧密连锁的SSR标记RM206进行分子标记辅助选择。通过分子标记检测、田间抗性鉴定和农艺性状选择,BC3F3株系中获得Xa23基因纯合且农艺性状稳定的培矮64S、明恢86和C418。获得的恢复系或不育系及配制的杂交组合在田间对我国7个以及菲律宾P1和P6白叶枯病生理小种都具有抗性。     

分子标记辅助培育水稻抗白叶枯病和稻瘟病三基因聚合系

水稻的白叶枯病和稻瘟病是水稻的两大主要病害, 通过分子标记辅助选择与传统的杂交、自交相结合的方法, 将抗稻瘟病的Pi9(t)基因和抗白叶枯病的Xa21及Xa23基因聚合到同一株系中, 经多代大田或/和温室接菌鉴定、室内标记选择和田间农艺性状的筛选, 获得了4个三基因聚合且农艺性状优良的株系L17~L20。用不同国家和地区的20个稻瘟病小种、中国流行的7个白叶枯病菌系C1~C7以及安徽省流行的白叶枯病菌系进行大田或/和温室抗病性鉴定, 结果显示, 株系L17~L20对20个稻瘟病小种均表现出抗性, 抗性水平与Pi9(t)基因的供体亲本75-1-127相当, 抗谱相同; 对白叶枯病的抗性和抗谱与Xa23基因相似, 不论在苗期还是在成株期均抗白叶枯病。与Xa21、Xa23基因的供体亲本M12和CBB23相比, 成株期的抗性水平有所增强。利用多重PCR技术, 在同一PCR反应中可同时选择Pi9(t)和Xa21基因, 提高了PCR选择效率。     

利用分子标记辅助选择聚合Pi9(t)和Xa23基因

利用分子标记辅助选择将广谱高抗稻瘟病的Pi9(t)基因和全生育期高抗白叶枯病的Xa23基因聚合到同一优良株系中,获得了含双基因的优良株系L10～L13.用来自不同地区的20个稻瘟病小种和中国流行的7个白叶枯病小种及安徽白叶枯病小种对聚合株系进行接菌鉴定,结果显示:聚合Pi9(t)和Xa23基因的株系L10～L13同时抗稻瘟病和白叶枯病;与稻瘟病的供体亲本75-1-127相比,抗性水平相当,均达抗级(R)水平,且抗谱相同;与白叶枯病的供体CBB23相比,对白叶枯病的抗性时期一致,均表现为全生育期抗性,而且抗性水平和抗谱相似.通过田间农艺性状的筛选,获得的双基因聚合系具有较好的农艺性状,可直接应用于生产或作为抗性亲本.     

蜀恢527抗水稻白叶枯病改良系的遗传背景和农艺性状分析

利用336个SSR标记和100个RAPD引物,对分子标记辅助选择获得的10个蜀恢527抗水稻白叶枯病改良系的遗传背景、白叶枯病的抗性和抗谱及农艺性状进行了研究。结果表明,10个改良株系在Xa21和Xa4位点纯合,抗我国7个病原型代表菌系CⅠ～CⅦ和菲律宾小种1和6（P1和P6）。在遗传背景分析检测的位点中,改良系与蜀恢527表现差异的SSR     

Improving bacterial blight resistance of'6078', an elite restorer line of hybrid rice, by molecular marker-assisted selection

Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (X00), is one of the most devastating diseases of rice world‐wide; it is also a serious problem of hybrid rice production in China. In this study, a molecular marker‐assisted introgression of Xa21, a gene highly resistant to a broad spectrum of Xoo strains, from ‘IRBB21’ was performed to improve the BB resistance of‘6078′, a new restorer line with high yielding potential. The entire process took one generation of crossing followed by three generations of backcrossing and one generation of selfing. The presence of Xa21 in each generation was determined by both polymerase chain reaction (PCR) and pathogen inoculation. Recombinations between Xa21 and flanking markers were identified by PCR analysis. Background selection was conducted in BC₁F₁ and BC₂F₁ using amplified fragment length polymorphism (AFLP) markers detecting a total of 129 polymorphic bands between‘6078’ and ‘IRBB21′. The individual selected in BC₃F₂, or‘6078′(Xa21), carried a fragment of less than 3.8 cM from the donor line in the Xa21 region on chromosome 11, and about 98.8% of the genetic background from the recurrent parent. The results showed that‘6078′(Xa21) had the same level and spectrum of BB resistance as the donor parent ‘IRBB21′, while maintaining the agronomic performance and combining ability of the original 6078. A significant increase in BB resistance was also achieved in the hybrid using 6078(Xa21) as the restorer line.     

水稻抗白叶枯病基因的聚合育种

白叶枯病是严重危害水稻生产的重要病害之一,利用聚合多个抗性基因的水稻品种是控制该病害的有效途径。本研究以携带Xa4、xa5、xa13、Xa214个抗白叶枯病基因的IRBB60与8个水稻新品系,组配8个杂交组合.应用分子标记辅助选择技术从F2和F3群体中共获得216个携带4个抗白叶枯病基因的纯合体。用华南地区的5个白叶枯病病原菌小种进行田间接种的试验表明,4个抗性基因聚合系的抗性水平高于只带1个抗性基因的近等基因系。这些聚合系可用作华南地区高抗白叶枯病水稻品种育种的亲本材料。     

超级杂交稻两优培九及其亲本生育后期的光抑制和早衰特性

以超级杂交稻两优培九（培矮64S/9311）和母本培矮64S（PA64S）以及父本中籼9311为材料，研究亲本和后代在自然条件下叶绿素荧光和活性氧代谢的差异。结果表明：强光高温下，在叶片衰老过程中，两优培九的叶绿素、蛋白质含量、PSⅡ原初光化学效率（Fv/Fm）和光化学猝灭系数（qP）减少较少，非光化学猝灭系数（qN）增加不明显     

Multiplex PCR genotyping for five bacterial blight resistance genes applied to marker‐assisted selection in rice (Oryza sativa)

Bacterial blight (BB) is the most economically damaging disease of rice in Asia and other parts of the world. In this study, a multiplex PCR genotyping method was developed to simultaneously identify genotypes of five BB resistance genes, Xa4, xa5, Xa7, xa13 and Xa21. The resistance R alleles were amplified using five functional markers (FMs) to generate amplicons of 217, 103, 179, 381 and 595 bp in IRBB66. Amplicons of 198, 107, 87, 391 and 467 bp corresponded to susceptible alleles in Taiwanese japonica rice cultivars. In backcross breeding programmes, the multiplex PCR assay was integrated into selection from a population using BB resistance donor IRBB66 crossed to rice cultivar ‘Tainung82’. Two plants with homozygosity for Xa4, xa5, Xa7, xa13 and Xa21 were selected from 1100 BC₂F₂ plants. In addition, the five BB resistance genes were also accurately identified in F₂ populations. This multiplex PCR method provides a rapid and efficient method for detecting various BB resistance genes, which will assist in pyramiding genes to improve durability of BB resistance in Taiwanese elite rice cultivars.     

Integrating marker assisted background analysis with foreground selection for identification of superior bacterial blight resistant recombinants in Basmati rice

Basmati rice is highly susceptible to bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae. Transfer of BB resistance genes from non‐Basmati sources to Basmati through cross‐hybridization requires strict monitoring for recovery of the desirable Basmati quality traits in the recombinants, which show complex inheritance pattern. We integrated background analysis using mapped microsatellite markers with foreground selection to identify superior lines that combine useful genes from a non‐Basmati BB resistance donor line IRBB55 with grain and cooking quality characteristics of the popular Basmati rice variety ‘Pusa Basmati 1’ (PB 1) employing backcross pedigree strategy. Foreground selection using linked markers ensured presence of two genes, xa13 and Xa21 for BB resistance from IRBB55, and the recurrent parent PB 1 allele for the waxy locus giving intermediate amylose content and maintainer allele at fertility restorer locus in the BC₁F₅ recombinants. Background analysis enabled selection of recombinants with recurrent parent genome to the extent of 86.3% along with the quality traits. The extent of introgression of non‐Basmati donor chromosome segments in the superior selections was estimated to be < 7.8 Mb and < 6.7 Mb in the xa13 and Xa21 linked genomic regions, respectively. Association mapping identified three quantitative trait loci, one each for 1000‐grain weight, fertile grains/panicle and cooked kernel length. The backcross‐pedigree breeding strategy facilitated recovery of additional desirable characteristics from the donor in some of the selections. The elite selection Pusa 1460‐01‐32‐6‐7‐67 with maximum genomic background and quality characteristics of the recurrent Basmati parent gave resistance reaction against BB, similar to that of the non‐Basmati resistant check variety and recorded an yield advantage of 11.9% over the best check in the multiplication agronomic trial in the Basmati growing region of India. This line, which has been released as a new variety in the name of ‘Improved Pusa Basmati 1’ for commercial cultivation in India, is an example of successful application of marker assisted selection to variety development.     

水稻抗除草剂基因bar的转育研究

本研究以转bar基因水稻"HR"为供体,两系法亚种间杂交稻优势组合"两优培九"的恢复系"9311"为受体,经过3次回交转育成了对Basta除草剂具有抗性的9311近等基因系"9311HR",经PCR分析确认已导入bar基因.比较试验表明,9311HR及其与"培矮64S"配制的杂种的产量、米质、抗性等主要农艺性状分别与9311和两优培九十分相似.表明通过回交将     

Marker assisted introgression of bacterial blight resistance in Samba Mahsuri,an elite indica rice variety

Samba Mahsuri (BPT5204) is a medium slender grain indica rice variety that is very popular with farmers and consumers across India because of its high yield and excellent cooking quality. However, the variety is susceptible to several diseases and pests, including bacterial blight (BB). We have used PCR based molecular markers in a backcross-breeding program to introgress three major BB resistance genes (Xa21, xa13 and xa5) into Samba Mahsuri from a donor line (SS1113) in which all the three genes are present in a homozygous condition. At each backcross generation, markers closely linked to the three genes were used to select plants possessing these resistance genes (foreground selection) and microsatellite markers polymorphic between donor and recurrent parent were used to select plants that have maximum contribution from the recurrent parent genome (background selection). A selected BC₄F₁ plant was selfed to generate homozygous BC₄F₂ plants with different combinations of BB resistance genes. The three-gene pyramid and two-gene pyramid lines exhibited high levels of resistance against the BB pathogen. Under conditions of BB infection, the three-gene pyramid lines exhibited a significant yield advantage over Samba Mahsuri. Most importantly, these lines retain the excellent grain and cooking qualities of Samba Mahsuri without compromising the yield as determined in multi-location trials. This work demonstrates the successful application of marker-assisted selection for targeted introgression of multiple resistance genes into a premium quality rice variety. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. R. M. Sundaram and M. R. Vishnupriya have contributed equally to this work.