通过生育期基因型选择避免稻瘟病抗性与结实率的遗传累赘

利用重组自交系研究表明,在水稻第6染色体短臂上稻瘟病抗性基因Pi25(t)与控制结实率和每穗实粒数的QTL之间存在遗传累赘。为了验证这种关系,采用了更大的遗传群体进行分析,结果表明稻瘟病抗性与结实率存在遗传累赘,但未检测到稻瘟病抗性与每穗实粒数存在遗传累赘。通过对第7染色体长臂RM2-RM214区间抽穗期基因（qHD 7）型背景进行选择,可以打破或避免稻瘟病抗性与结实率的遗传累赘。为了进一步验证这种关系,选择Pi25(t)区间基因型不同、RM2-RM214区间基因型相同、其他染色体区间基本一致的两个株系发展新群体进行分析,除第6染色体的结实率QTL可以分解成2个效应较小的QTL（qSF 6 1和qSF 6 2）外,当第7染色体RM2-RM214区间基因型为中156背景时,Pi25(t)与结实率QTL（qSF 6 2）存在遗传累赘,且qSF 6 2来自父本谷梅2号的等位基因起减效作用;当第7染色体RM2-RM214区间基因型为谷梅2号背景时,第6染色体上没有检测到结实率QTL。上述结果说明在特定育种材料中对抽穗期基因进行选择可以成功打破或避免稻瘟病抗性与结实率的遗传累赘,为水稻以及其他作物的高产抗病育种提供了一种新途径。     

抗稻瘟病水稻材料谷梅2号中主效抗稻瘟病基因的成簇分布

应用由籼稻组合中156/谷梅2号衍生的304个重组自交系,构建了由177个标记组成、覆盖12条水稻染色体的连锁图谱,定位控制水稻稻瘟病抗性的主效基因。前期定位的控制对中国稻瘟菌菌系92 183（小种ZC15）穗瘟抗性的基因Pi25(t)的位置得到进一步确认,位于第6染色体标记A7和RG456之间,与A7和RG456的遗传距离分别为1.7和15 cM;发现群体对菲律宾稻瘟菌菌系Ca89（4谱系）的叶瘟抗性由单基因控制,将该暂命名为Pi26(t)的基因定位于第6染色体标记B10和R674之间,与B10和R674的遗传距离分别为5.7和25.8 cM。两个基因座位上的抗病等位基因均来源于谷梅2号,表明谷梅2号中存在控制水稻稻瘟病抗性的基因簇。     

具抗稻瘟病基因Pi25杂交稻恢复系的分子标记辅助选育

 从组合中156/谷梅2号所衍生的重组自交系群体中选择携带抗稻瘟病基因Pi25的3个株系,分别与高产恢复系9308和3 11配组,通过单粒传法获得的6个F6重组自交系群体用于研究。在Pi25先前定位的遗传图谱上,新增加与该基因紧密连锁的分子标记RM3330和A7,并结合RM3330和A7分子标记辅助选择的结果,共筛选到了109个Pi25基因纯合的株系。用现有的与恢复基因连锁的标记对这109个株系进行二次筛选,最终获得20个Pi25基因和恢复基因均纯合的株系。对育性恢复力鉴定试验表明,选育出的抗病恢复系具有较好的恢复能力,并已应用于育种实践;人工稻瘟病接种试验证实,所用标记不同,分子标记辅助选择的效率差异显著,单个标记辅助选择符合率均不高,而采用目标基因两侧连锁的标记同时进行辅助选择,则可以明显提高分子标记辅助选择符合率。     

Clustering of Major Genes Conferring Blast Resistance in a Durable Resistance Rice Cultivar Gumei 2

By using 304 recombinant inbred lines derived from indica rice cross Zhong 156/Gumei 2, a linkage map consisting of 177 marker loci and covering 12 rice chromosomes was constructed and employed for mapping genes conferring blast resistance in rice. Genomic location of gene Pi25(t) conferring neck blast resistance to the Chinese isolate 92-183 (race ZC15) was verified to be located between markers A7 and RG456 on chromosome 6, with genetic distances of 1.7 cM and 1.5 cM to A7 and RG456, respectively. Leaf blast resistance of Gumei 2 to the Philippine isolate Ca89 (lineage 4) was found to be controlled by a single gene. The gene tentatively designated as Pi26(\) was located between makers B10 and R674 on chromosome 6, with genetic distances of 5.7 cM and 25.8 cM to B10 and R674 respectively. Resistant alleles at both gene loci were derived from Gumei 2, indicating an existence of resistance gene cluster in Gumei 2.     

Clustering of Major Genes Conferring Blast Resistance in a Durable Resistance Rice Cultivar Gumei 2

By using 304 recombinant inbred lines derived from indica rice cross Zhong 156/Gumei 2, a linkage map consisting of 177 marker loci and covering 12 rice chromosomes was constructed and employed for mapping genes conferring blast resistance in rice. Genomic location of gene Pi25(t) conferring neck blast resistance to the Chinese isolate 92-183 (race ZC15) was verified to be located between markers A7 and RG456 on chromosome 6, with genetic distances of 1.7 cM and 1.5 cM to A7 and RG456,respectively. Leaf blast resistance of Gumei 2 to the Philippine isolate Ca89 (lineage 4) was found to be controlled by a single gene. The gene tentatively designated as Pi26(t) was located between makers B10 and R674 on chromosome 6, with genetic distances of 5.7 cM and 25.8 cM to B10 and R674 respectively. Resistant alleles at both gene loci were derived from Gumei 2,indicating an existence of resistance gene cluster in Gumei 2.     

Pigm特异性选择标记的开发及其在粳稻穗颈瘟抗性育种中的利用

【目的】Pigm是一个广谱的稻瘟病抗性基因,源自持久抗性品种谷梅4号,与Piz、Piz-t、Pi2、Pi9和Pi40等互为复等位基因但抗谱存在差异。为了更好地在分子辅助选择育种中利用Pigm基因,开发与Pigm特异性标记具有重要意义。【方法】在已有文献报道定位结果的基础上,通过随机测序获得了一段谷梅4号基因组的特异序列,并据此开发了一组用于筛选Pigm基因的分子标记,进一步选取江淮稻区3个不同生态型代表性粳稻品种作为受体,利用分子标记辅助选择结合对抗性基因的背景检测将Pigm基因导入受体品种。【结果】Pigm-4标记位于Pigm基因簇内部,与抗病功能元件PigmR紧密连锁,对不同类型的品种检测发现该标记特异性强,且利用该标记可将Pigm与Piz、Piz-t、Pi2、Pi9以及Pi40区分开来。对受体亲本稻瘟病抗性基因的检测和接种结果分析发现江淮稻区粳稻品种虽然携带了Pib、Pi54、Pita、Pb1中的2~3个基因,但是对强毒力的稻瘟病小种抗性普遍不强,而3种代表性粳稻背景下导入Pigm基因均可显著提高其对穗颈瘟的抗性水平。【结论】Pigm可以作为抗稻瘟病粳稻育种的有利基因资源加以利用,而Pigm-4是分子标记辅助筛选Pigm的优异标记。     

应用候选基因定位水稻抗稻瘟病QTL

 应用经克隆了的已知功能或有潜在功能的DNA序列,即候选基因，作为分子标记，在中156/谷梅2号F8重组自交系群体中进行水稻抗稻瘟病QTL的分析。大部分候选基因在水稻染色体上成簇分布，并且位于已知抗病基因簇区域。应用复合区间法检测到1个调控病斑大小和1个调控病斑数量的QTL，前者位于第1染色体CG36a~RM212区间，贡献率为4.17%，抗性等位基因来自父本谷梅2号；后者定位于第2染色体CG18a~RM263区间，贡献率为6.25%，抗性等位基因来自母本中156。同时检测到2对控制病叶面积和1对控制病斑大小的基因互作。这些QTL和互作基因涉及抗性基因同源序列、离子通道调控子以及编码致病相关蛋白和几丁质酶的基因，表明候选基因的应用有助于揭示QTL的功能。玉米锈病抗性基因Rp1与稻瘟病抗性有关，提示了利用水稻这个模式作物来克隆较大基因组中有利基因的可能性。     

Improvement of Upland Rice Variety by Pyramiding Drought Tolerance QTL with Two Major Blast Resistance Genes for Sustainable Rice Production

Varalu is an early maturing rice variety widely grown in the rainfed ecosystem preferred for its grain type and cooking quality. However, the yield of Varalu is substantially low since it is being affected by reproductive drought stress along with the blast disease. The genetic improvement of Varalu was done by introgressing a major yield QTL, qDTY_12.1, along with two major blast resistance genes i.e. Pi54 and Pi1 through marker-assisted backcross breeding. Both traits were transferred till BC₂ generation and intercrossing was followed to pyramid the two traits. Stringent foreground selection was carried out using linked markers as well as peak markers (RM28099, RM28130, RM511 and RM28163) for the targeted QTL (qDTY_12.1), RM206 for Pi54 and RM224 for Pi1. Extensive background selection was done using genome-wide SSR markers. Six best lines (MSM-36, MSM-49, MSM-53, MSM-57, MSM-60 and MSM-63) having qDTY_12.1 and two blast resistance genes in homozygous condition with recurrent parent genome of 95.0%-96.5% having minimal linkage drag of about 0.1 to 0.7 Mb were identified. These lines showed yield advantage under drought stress as well as irrigated conditions. MSM-36 showed better performance in the national coordinated trials conducted across India, which indicated that improved lines of Varalu expected to replace Varalu and may have an important role in sustaining rice production. The present study demonstrated the successful marker-assisted pyramiding strategy for introgression of genes/QTLs conferring biotic stress resistance and yield under abiotic stress in rice.     

稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用

【目的】来自籼稻品种谷梅4号的Pigm是一个对稻瘟病菌具有广谱和持久抗性的重要基因。为有效提高Pigm基因的选择效率,有必要开发具有特异性的共显性分子标记进行辅助育种。【方法】本研究根据谷梅4号Pigm位点存在的特异性核苷酸变异,利用Tetra-primer ARMS-PCR和KASP两种不同的基因分型技术开发出分子标记T-Pigm和K-Pigm,对不同品种(品系)以及淮稻9号/谷梅4号的F₂分离群体进行基因型检测,并结合穗颈瘟人工接种鉴定,对标记的准确性进行评价。【结果】序列比对分析表明,谷梅4号Pigm位点中Pigm-Nbs2基因起始密码子上游515 bp处存在一个特殊的单核苷酸变异。利用已开发的两种类型的分子标记能够有效区分3种不同的基因型,且基因型与穗颈瘟人工接种鉴定结果完全一致。【结论】利用分子标记T-Pigm和K-Pigm可以实现对Pigm基因型快速、准确的检测,加快抗稻瘟病水稻新品种的选育进程。     

稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用

【目的】来自籼稻品种谷梅4号的Pigm是一个对稻瘟病菌具有广谱和持久抗性的重要基因。为有效提高Pigm基因的选择效率，有必要开发具有特异性的共显性分子标记进行辅助育种。【方法】本研究根据谷梅4号Pigm位点存在的特异性核苷酸变异，利用Tetra-primer ARMS-PCR和KASP两种不同的基因分型技术开发出分子标记T-Pigm和K-Pigm，对不同品种(品系)以及淮稻9号/谷梅4号的F2分离群体进行基因型检测，并结合穗颈瘟人工接种鉴定，对标记的准确性进行评价。【结果】序列比对分析表明，谷梅4号Pigm位点中Pigm-Nbs2基因起始密码子上游515 bp处存在一个特殊的单核苷酸变异。利用已开发的两种类型的分子标记能够有效区分3种不同的基因型，且基因型与穗颈瘟人工接种鉴定结果完全一致。【结论】利用分子标记T-Pigm和K-Pigm可以实现对Pigm基因型快速、准确的检测，加快抗稻瘟病水稻新品种的选育进程。     

水稻白化转绿和穗顶端退化突变体vpa1的遗传分析和基因定位

目的 通过对水稻转绿和穗顶端退化等突变体的研究,可以鉴定更多与叶绿体发育和穗发育相关的基因。方法 在常规种植条件下比较突变体vpa1（virescent and panicle abortion 1)表型及主要农艺性状差异,利用分离群体分析和图位克隆法进行相关基因定位。结果 突变体vpa1表现苗期白化,并逐渐转绿恢复成正常叶色,抽穗后可明显观查到穗顶端退化表型。vpa1的主要农艺性状除了结实率以外,株高、穗长、每穗实粒数等均较野生型显著下降。遗传分析表明白化转绿和穗顶端退化表型受独立的两个隐性基因控制。控制白化转绿叶性状的Osv16定位于第3染色体RM3441和RM3029之间约125kb物理区间内,区间内未见白化转绿性状相关基因的报道。控制穗顶端退化性状的Ospaa10定位于第1染色体RM11157和RM5972之间,区间内物理距离约190kb,区间内未见穗顶端退化相关基因的报道。结论 Osv16和Ospaa10两个基因的突变导致vpa1的叶色和穗型同时出现变异,为白化转绿基因Osv16和穗顶端退化基因Ospaa10的克隆和功能研究打下了基础。     

QTL analysis of panicle morphology response to irrigation regime in aerobic rice culture

In water-efficient rice production, grain yield is often constrained by panicle size. The objective of this study was to genetically dissect the response of panicle morphology to irrigation regimes in aerobic rice culture. We grew ‘Akihikari’ (a lowland japonica cultivar) × ‘IRAT109’ (an upland japonica cultivar) backcross inbred lines in aerobic soils with full or limited irrigation for 2 years, and examined 4 panicle traits—number of florets per panicle (FPP), number of primary branches per panicle (BPP), number of florets per primary branch (FPB), and frequency of pre-flowering floret abortion (%FA)—and grain yield. QTLs for BPP were detected in both the irrigation regimes but QTLs for FPB and %FA were detected mostly only in either of the irrigation regimes. The QTL for FPP on chromosome 2 (RM3421–RM213) coincided with that for yield under full irrigation, showing that this QTL is related to sink capacity and yield potential in aerobic rice culture. On the other hand, the QTL for FPB on chromosome 1 (RM3148–RM243) coincided with that for yield under limited irrigation, when water deficit was moderate. The QTL for root axis length at vegetative stage, previously identified in the same mapping population, was located near this region. This study unravelled the complicated genetic control on panicle morphology in aerobic rice culture, and suggested the positive roles of the dehydration avoidance mechanism by vigorous root growth on panicle size and yield under dry soil conditions.     

应用剩余杂合体衍生的近等基因系分解水稻产量性状QTL

以杂合区间为RM587-RM402的水稻剩余杂合体（RHL）衍生群体为材料,应用SSR标记检测,筛选到杂合区间分别为RM587-RM225、RM204-RM6119和RM6119-RM402的3个单株,进一步检测其F2群体,分别获得母本纯合型材料10株、父本纯合型材料10株和杂合型材料20株。种植这3套近等基因系材料,考查单株产量及其构成因子每株穗数、每穗实粒数和千粒重。经应用目标区间内等位基因效应分析和交迭重组染色体片段代换系分析,分解出3个控制每穗实粒数的QTL和2个控制单株产量的QTL,这些QTL分别位于物理距离为0.66 ~ 2.49 Mb的区间中,全部表现为加性作用为主,增效等位基因除qNFGP6 1来自父本密阳46外,其余均来自母本珍汕97B。提出了构建新型遗传材料,提高水稻QTL精细定位效率的策略。     

应用RFLP标记研究水稻的广亲和基因

 选用45个分布于10条染色体上的DNA探针，结合4种限制性内切酶，检测了广亲和品种Pecos和测验种南京11、秋光之间DNA限制性片段长度多态性(RFLP)。发现其中15个探针能在Pecos、南京11和秋光之间表现多态性。进一步选用14个表现多态性的探针和色素原基因C分析了三交群体（Pecos／南京11//秋光）中单株结实率的分离。结果表明第6染色体上的RFLP标记RG138、RG64 RG456以及色素原基因C与籼粳杂交亲和性有显著的连锁关系；位于第12染色体上的标记RG81也与三交群体单株结实率显著相关，根据标记与性状的共分离资料，初步绘制了第6染色体上该籼粳杂交亲和性座位的RFLP连锁图。此外。还发现第12染色体上的两个标记RG81和RG323在三交群体中两种基因型的分离比例不符合1∶1。     

太湖流域粳稻地方品种黑壳子粳抗稻瘟病基因的分子定位

以广谱、高抗稻瘟病的太湖流域粳稻地方品种黑壳子粳与感病品种苏御糯杂交,产生F1、F2、F2∶3及F5∶6重组自交系群体,用日本稻瘟病鉴别菌系北1接种鉴定。黑壳子粳对北1的抗性是由1对显性主效基因控制的,定名为Pi hk1(t) 。根据不同杂交世代群体对北1的抗、感反应,结合SSR分子标记,将黑壳子粳中的Pi hk1(t) 基因定位在水稻第11染色体长臂末端,与RM7654和RM27381两个标记的遗传距离分别为0.9 cM和1.6 cM。     

水稻小穗簇生基因OsCL6的定位及候选基因分析

【目的】通过对水稻小穗簇生基因的定位与候选基因分析,为进一步克隆幼穗发育过程中缩短枝梗长度造成小穗簇生的功能基因奠定基础。【方法】以航天诱变育种技术处理常规优质稻品种粤农丝苗获得一个稳定遗传的小穗簇生突变体cl6为试验材料,与粤金银占构建F₁、F₂作图群体,对簇生基因进行定位,结合转录组测序对候选基因进行预测与评价。【结果】遗传分析结果表明,突变体cl6的簇生性状由非完全显性单基因控制,混合分组分析法将该基因定位于第6染色体上约416 kb的区间。进一步通过表达谱分析、转录组测序技术、基因序列差异性分析和qRT-PCR验证等筛选出OsFBK16为最佳候选基因,其5′-UTR区域发生9个碱基的插入突变,暗示该基因可能通过二级结构改变调控转录或翻译水平,参与水稻幼穗发育过程中枝梗的分化。【结论】本研究结果为解析水稻二次枝梗和小穗梗缩短机制奠定一定理论基础。     

Genetic bases of source-,sink-,and yield-related traits revealed by genome-wide association study in Xian rice

The source-sink relationship determines the ultimate grain yield.We investigated the genetic basis of the relationship between source and sink and yield potential in rice.In two environments,we identified quantitative trait loci(QTL)associated with sink capacity(total spikelet number per panicle and thousand-grain weight),source leaf(flag leaf length,flag leaf width and flag leaf area),source-sink relationship(total spikelet number to flag leaf area ratio)and yield-related traits(filled grain number per panicle,panicle number per plant,grain yield per plant,biomass per plant,and harvest index)by genome-wide association analysis using 272 Xian(indica)accessions.The panel showed substantial variation for all traits in the two environments and revealed complex phenotypic correlations.A total of 70 QTL influencing the 11 traits were identified using 469,377 high-quality SNP markers.Five QTL were detected consistently in four chromosomal regions in both environments.Five QTL clusters simultaneously affected source,sink,source–sink relationship,and grain yield traits,probably explaining the genetic basis of significant correlations of grain yield with source and sink traits.We selected 24 candidate genes in the four consistent QTL regions by identifying linkage disequilibrium(LD)blocks associated with significant SNPs and performing haplotype analysis.The genes included one cloned gene(NOG1)and three newly identified QTL(qHI6,qTGW7,and qFLA8).These results provide a theoretical basis for high-yield rice breeding by increasing and balancing source–sink relationships using marker-assisted selection.     

Specific Spikelet Fertility as an Indicator of Cold Tolerance Identification at Booting Stage in Rice

Spikelet fertility is an uppermost evaluation indicator of cold tolerance in rice at the booting stage. Though many evaluation indicators have adopted by scientists, the whole spikelet fertility (SF) is one of the most important evaluation indexes under low- temperature stress conditions [1-3], which has practical significance in rice production. However, spikelets at different developmental stages have different sensitivities to low-temperature. Moreover, even within the same panicle the spi…     

水稻品种8504的广亲和性及其利用价值

对原产澳大利亚的品种８５０４的广亲和性进行了测交验证，并分析和评价了该广亲和品种的利用价值．该品种广亲和性好、亲和谱广，熟期适宜，米质较好，中抗稻瘟病，与籼、粳杂交，杂种优势均较强，且广亲和性遗传简单．