野生稻（O.rufipogon）新胞质改良不育系稻米品质的研究

通过核置换回交，获得一种普通野生稻（O. rufipogon）雄性不育细胞质，称为FA细胞质，育成新质源不育系金农1A。研究表明，FA细胞质不育系与WA和HL不育系的恢保关系不同，野败型的保持系和恢复系、红莲型的保持系和恢复系都可以作为FA细胞质的保持系，而不能成为FA细胞质的恢复系。这是一种新发掘的水稻雄性不育细胞质源。FA细胞质突破了WA和HL细胞质恢保关系的遗传局限，扩大了保持系育种范围，可以用品质优良的栽培品种培育成优质稻米不育系。新质源不育系金农1A不育度高，不育性稳定，农艺性状优良，抗稻瘟病和白叶枯病，开花时间早，稻米12项品质指标全部达到农业部颁发的优质米一级或二级标准，实质性地提高了不育系的稻米品质和综合水平，为培育优质米杂交稻奠定良好的遗传基础。     

四种细胞质六千辛A粳稻不育系育种利用特性的比较研究

对由粳稻品系六千辛转育成的4种细胞质的一套同核异质不育系的不育性、开花习性、异交特性及可恢复性进行了鉴定。结果表明:(1)包台(BT)型、红莲(HL)型、茶野(CL)型六千辛A花粉以染败为主,WA型六千辛A花粉100%典败;BT型六千辛A平均自交结实率为0.60%,生产应用的安全性较差;HL型、CL型六千辛A无自交结实现象,不育性较BT型六千辛A稳定。(2)不育系的花粉败育程度影响倒一节间长度,花粉败育越彻底,倒一节间越短,从而使穗伸出度和株高相应缩短;因此,WA型六千辛A表现包颈,而BT型、HL型、CL型六千辛A均无包颈。(3)WA型六千辛A开花不集中,花时明显滞后,花期明显拉长,异交结实率低;HL型、CL型六千辛A的开花习性及异交结实率与BT型六千辛A无显著差异,明显好于WA型六千辛A。(4)BT型六千辛A的可恢复性最好,HL型、CL型六千辛A次之,WA型六千辛A最难恢复;从BT型粳稻恢复系中可以筛选到HL型或CL型粳稻不育系的恢复系。HL型、CL型粳稻不育系较好地协调了育性稳定性、开花习性和可恢复性几方面的矛盾。因此,选育HL型、CL型粳稻不育系及其相应的恢复系,可能是解决目前生产上杂交粳稻BT型不育系育性不够稳定及细胞质单一问题的突破口。     

野生稻（Oryza rufipogon）新质源雄性不育恢复系的研究

发掘野生稻（O. rufipogon）新型雄性不育细胞质源,育成新质源优质米不育系的基础上进一步研究新质源雄性不育恢复系的育种技术—FA型细胞质雄性不育恢复系定向育种。用野生稻（非轮回亲本）与籼稻品种明恢63（轮回亲本）杂交和多次回交,后代再经过自交,将野生稻中的可育基因分离、转移、重组、整合到明恢63遗传背景中,获得农艺性状似明恢63,花粉和小穗全可育不分离的野生稻新质源恢复系金恢1号。用新质源不育系与金恢1号组配两个组合,其花粉和小穗育性都恢复到正常可育水平,产量高,米质优,实现了新质源不育系三系配套应用和大幅度提高杂交稻稻米外观品质的目的。这项育种新技术可以将水稻可育基因（恢复基因）转移到任一水稻品种中育成细胞质雄性不育恢复系,突破了新质源恢复系育种的技术瓶颈,极大地提高了恢复系利用稻种资源的育种潜力,为FA型新质源优质米不育系的杂交稻育种开辟了一条崭新的途径。新型（FA）细胞质源杂交稻可能对丰富杂交稻细胞质遗传多样性、提高杂交稻亲本对稻种资源的利用潜力、以及实质性提高杂交稻的稻米品质和产量水平都将产生积极和深远的影响。     

水稻质核互作雄性不育系选育的反向杂交法研究

不同的部分保持系可能存在不同的微效恢复基因,通过有性杂交,产生基因重组,能够获得完全保持系类型,但只有在不育细胞质中才能观察得到微效恢复基因是否存在以及它们的作用大小.反向杂交法以不育细胞质为选择背景,在杂交后代植株中直接观察到微效恢复基因的表达,获得的完全不育株,在一定程度上排除微效恢复基因,不育株再通过高温处理转换为可育后自交,来自不育株的微效恢复基因可以进一步排除掉,从而产生出没有(或很少)微效恢复基因的"亲本",利用该"亲本"高温处理后仍可转换为可育的特性,作为父本进行杂交或回交育种,在其后代中获得没有微效恢复基因的完全保持系.该研究为Cp 26不育细胞质源创造出了完全保持系.如果在田间鉴定出优异的完全不育株,对其进行单倍体育种(诱导孤雌生殖或花培),选择到没有(或很少)微效恢复基因的纯合不育株.再对其进行花培,筛选可育的突变体;或者利用纯合不育株的原生质体与一个已破坏细胞核的可育系原生质体融合,都可能得到具有纯合不育株细胞核和可育细胞质的保持系,而能够完善和改进反向杂交法.反向杂交法不但能够为所谓不能保持的不育细胞质源创造出保持系,而且有利于加强新不育系选育的目标性和预见性,提高不育系配合力和培育不同类型优异不育系.     

水稻体细胞无性系雄性不育突变

对5个类型的水稻不育系进行了幼穗培养, 在红源A、 包源A和W6154s中, 共获得了10例雄 性不育变异株, 水稻花粉败育可分为无花粉、 典败、 圆败和染败四种类型。 发现了不育 花粉败育类型之间可以相互转换现象。 对雄性不育变异株用一批现有CMS不育系的保持系和恢复系进行测交和回交, 有的变异株其 恢保关系发生了变化,     

Development of a DNA marker for distinguishing CMS lines from fertile lines in rice (Oryza sativa L.)

The main objective of the present study was to identify mitochondrial DNA based marker, which can distinguish male sterile and fertile counterparts of the cytoplasmic male sterile (CMS) lines used in production of rice hybrids. Amplified fragment length polymorphism (AFLP) analysis in CMS lines: IR58025A & IR62829A and their respective maintainers: IR58025B & IR62829B identified a polymorphic DNA fragment of about 510 bp size that was present in both CMS (A) and absent in their maintainer (B) lines. Sequencing followed by database analysis of the polymorphic fragment indicated about 97% similarity with mitochondrial NADH gene subunits of rice, maize and wheat. Based on the variable sequence regions, a site specific primer pair (BF-STS-401) was designed. PCR analysis showed that BF-STS-401 could amplify a strong band of 464 bp size in CMS and a faint band of the same size in maintainer line. To act as a positive control and avoid possible errors in PCR, BF-STS-401 was multiplexed with a new primer pair (BF-STS-402), derived from mitochondrial atp9 subunit of rice, producing monomorphic amplification indiscriminately in both CMS and maintainer lines. Both the primer pairs in combination clearly differentiated CMS lines from their corresponding maintainer lines. This primer combination was validated in a set of diverse genotypes consisting of different sources of CMS lines, restorer lines, hybrids, varieties and mixed samples from private seed companies. Our results suggested that the multiplex primer pairs developed in this study can be effectively utilized to assess the genetic purity in commercial seed lots of CMS lines and hybrids of rice.     

水稻核不育系柱头性状的主基因+多基因遗传分析

柱头性状是影响水稻不育系异交繁殖和杂交水稻制种产量的重要性状。为创制长柱头、高外露率的水稻温敏核不育系提供遗传信息,调查了短柱头、低外露率的粳型光温敏核不育系7001S和长柱头、高外露率的温敏核不育系紫泰S及其杂交、自交获得的F_1、F_2群体(350个株系)和F_2:3群体(320个株系)的4个柱头性状,分析了4个性状之间的相关性,并运用主基因+多基因混合遗传模型,对2个世代4个性状进行了遗传分析。结果表明,4个柱头性状间均表现出极显著正相关,相关系数介于0.262和0.895之间。柱头长度、花柱长度、柱头和花柱总长度(以下简称柱花总长度)均表现出受2对主效基因和微效基因共同控制,除F_2群体中柱花总长度的2对主基因表现为等加性效应和等显性效应外,其余均表现为加性-显性-上位性效应,3个性状均表现出以主基因间的上位性效应为主;F_2群体柱头外露率受2对加性-显性-上位性主基因+多基因控制,而F_2:3群体则表现为受1对加性-显性主基因+多基因控制,以主基因间的加性效应为主。2个世代中的4个柱头性状均以主基因遗传为主。     

Influence of Male Sterile Cytoplasms on the Physico-Chemical Grain Quality Traits in Hybrid Rice (Oryza Sativa L.)

Summary Possible unfavourable influence of sterility inducing cytoplasm on physico-chemical grain quality traits in rice hybrids is one of the important concerns hindering the large-scale adoption of hybrid rice technology. Produce from 23 pairs of CMS line × restorer (AF₁) and maintainer line × restorer (BF₁) cross combinations carrying different cyto-sterile sources (WA, ARC, Mutagenized IR 62829B and Kalinga I) was compared for various grain and cooking quality traits. The milling recovery in rice hybrids was not influenced by the sterile cytoplasm. For kernel dimensions before and after cooking there were both favourable and unfavourable cytoplasmic effects, which varied in magnitude depending upon the sterile cytoplasm and parental combinations. Similar results were obtained for kernel elongation and gelatinization temperature. The most widely used WA cytoplasm had minimum instances of unfavourable influence. In general, the cytoplasmic influence was found to be highly cross-specific and depended on the nuclear background of CMS line and fertility restorer. Availability of alloplasmic CMS lines carrying different cyto-sterile sources in the same nuclear background would help in ascertaining the cytoplasmic influence in a more comprehensive manner.     

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     

水稻强分蘖两系不育系‘明S’农艺性状研究

为深入研究水稻两系核不育系‘明S’特性,以期配制高产组合,在生产上得到广泛应用。以‘培矮64S’为对照,相同栽培管理条件下,对‘明S’与‘培矮64S’的植物学特征和生物学特性进行分析。结果表明‘明S’具有株叶形态好、分蘖力强、成穗率高、稻瘟病抗性好等特点。单株有效穗数17个,花粉育性镜检,为无花粉型和少数典败单株,不育率100%,不育度100%,自交结实率为0。柱头外露率高,为84.7%,异交结实率70%以上。稻米品质较好,糙米率、整精米率、胶稠度、碱消值达到一级标准;抗稻瘟病、白叶枯病,纹枯病发病较轻。可见两系不育系‘明S’是一个具有强分蘖特性并且抗稻瘟病的温敏核不育系。     

Assessment of purity of rice CMS lines using cpDNA marker

AFLP technique was used to analyse the polymorphism between rice cytoplasmic male sterility (CMS) line Jin2A and its maintainer Jin2B. A stable differential band was discovered, and sequence analysis showed that Jin2A contained a more tandem repeat of 6 base pairs (AGAAAA) than Jin2B. Further studies confirmed that the diversity came from cpDNA and occurred at three kinds of abortive cytoplasmic genotypes. Accordingly, specific primers were designed and utilized to assess the purity of rice CMS lines during multiplication with pollen fertility and seed setting rates of bagged panicles as control. The result indicated that this cpDNA locus could be utilized to precisely distinguish maintainer plants from rice CMS lines. PCR analysis was consistent with that from Grow-out test in CMS line seed purity assessment during multiplication, despite it was helpless in distinguishing F₁ hybrids from CMS lines due to similar cytoplasms. Because of fewer hybrid and more maintainer off-plants, this cpDNA locus was still appropriate for seed purity assessment of rice CMS line during multiplication. This is first report that a marker on cpDNA could be utilized to assess the genetic purity of rice CMS lines with three abortive cytoplasmic genotypes.     

同核异质滇I型粳稻不育系及其保持系的光合特性

滇I型不育系和保持系是我国粳稻杂种优势利用最主要的技术体系之一。本研究利用Li-6400便携式光合测定系统测定了79对滇I型同核异质粳稻不育系及保持系上三叶的净光合速率(P_n)、气孔导度(G_s)、胞间CO₂ 浓度(C_i)、蒸腾速率(T_r)、水分利用效率(WUE)、气孔限制值(L_s)和叶绿素相对含量(SPAD)。结果表明,不育系间及保持系间的光合参数在3个叶位上都达到了极显著水平,基因型间的这种显著差异为高光效育种提供了宝贵的遗传基础。同一叶位上不育系与保持系间的光合参数具有显著的正相关。除SPAD值外,其他光合参数值在不育系与保持系间没有显著差异,但不育系的SPAD值显著大于保持系。叶绿素相对含量不是粳稻提高光合作用的限制因子;滇I型细胞质对光合作用没有负效应;利用滇I型细胞质转育的不育系的叶绿素含量很可能有所提高。基于剑叶光合参数值对保持系及不育系的聚类,不育系和保持系可分别分为高和较低光合速率的类型。本研究为滇I型不育系的高光效育种提供了理论基础,也可作为杂交粳稻高光效的理论参考。     

杂交稻抗白叶枯病的遗传机制

8种不同胞质类型的不育系及其保持系与9个恢复系按P×Q交配设计分别配制不育系杂种和保持系杂种, 接种白叶枯病(Xoo)“强”致病力菌系ZHE-173和“弱”致病力菌系KS-6-6, 研究杂交稻抗白叶枯病的遗传机制。 结果表明： (1) 不育系或保持系、 恢复系以及不育系或保持系与恢复系间的互作对不育系杂种或保持系杂种抗性的影响均     

乙烯与水稻细胞质雄性不育的关系

从幼穗发育的IV到VII期,水稻细胞质雄性不育系(珍汕97A)幼穗和叶片的ACC含量和乙烯释放速率均高于其保持系(珍汕97B)。外施乙烯释放剂乙烯利使保持系花粉可育度明显下降;外施ACC合成酶抑制剂AVG引起两系幼穗ACC含量和乙烯释放速率下降,并使不育系花粉育性得以部分恢复,而在外施AVG的同时再施以乙烯利则AVG的恢复作用消失。     

Identification of restorers and maintainers for developing basmati and non-basmati hybrids in rice, Oryza sativa

Twenty‐seven improved aromatic lines of germplasm and 18 non‐aromatic disease‐resistant genotypes of rice were test‐crossed with four cytoplasmic male‐sterile lines (IR 58025A, IR 62829A, PMS 3A and PMS 10A). Thirteen aromatic and 10 non‐aromatic genotypes were selected based on pollen fertility, and crosses were repeated to confirm sterility‐maintaining and fertility‐restoring ability. Genotypes were categorized as effective restorers (> 80% spikelet fertility), partial restorers (21‐79% spikelet fertility) and maintainers (< 1% spikelet fertility). The effective basmati restorers identified were Basmati 385, Chandan, P1031‐8‐5‐1, HKR 241‐IET‐12020, SAF Khalsa 7 and Karnal Local. The basmati maintainers identified were Basmati 370, Pusa basmati 1, P615‐K‐167‐13 and P1173‐4‐1. The frequency of restorers obtained was higher for the non‐aromatic than the aromatic basmati type. The performance of restorers varied with cytoplasmic male‐sterile (CMS) line, location and season of testing. The differential ability to restore fertility in the CMS lines that have the wild abortive (WA) cytosterile system could result from different nuclear backgrounds of the CMS lines. These restorers and maintainers possess acceptable grain dimensions, a desirable degree of aroma, volume expansion through linear kernel elongation and cooking quality characteristics of basmati rice. These genotypes will contribute to developing basmati hybrids and provide restorers and maintainers with acceptable key basmati quality characteristics.     

胞质雄性不育和核雄性不育辣椒内源激素含量的变化

采用酶联免疫吸附法(ELISA)测定和比较分析了CMS和GMS辣椒不育系(株)和相应保持系(可育株)花蕾和叶片中的内源IAA,Z ZR,GA3,ABA含量及比值的变化。结果表明,CMS雄性不育系和保持系花蕾中IAA,Z ZR,GA3和ABA含量的变化趋势与GMS不育株和可育株间变化趋势基本相同,即不育系(株)IAA,Z ZR,GA3和ABA含量低于保持系(可育株);叶片中Z ZR,GA3和ABA含量变化趋势GMS和CMS也基本一致,即不育系(株)中的含量高于保持系(可育株);花蕾中IAA/ABA,(Z ZR)/ABA,GA3/ABA激素比例的变化GMS与CMS也相同,即不育系(株)比值小于保持系(可育株)。说明虽然GMS和CMS辣椒控制雄性不育的基因不同,但在调节雄性不育时激素的变化规律大多相同,辣椒雄性不育在生理生化机理上可能存在一些相同的特点。     

红莲型水稻不育系和保持系线粒体基因组BAC文库的构建

以红莲型(HL)细胞质雄性不育系和保持系为材料, 构建了水稻线粒体基因组的BAC文库. 每个文库保存约2300个菌落, 外源插入片段介于9～25 kb之间. 以线粒体基因为探针对文库进行菌落原位杂交验证, 均筛选到了阳性克隆. 构建的两个文库为进一步研究水稻线粒体基因组的结构特点, 为克隆与红莲型水稻细胞质雄性不育相关的线粒体基     

Rf-1位点CAPS标记对水稻不同胞质雄性不育育性恢复系的关系分析

水稻育性恢复基因Rf-1是目前水稻上克隆的唯一恢复基因.通过位于Rf-1位点的两个特异性CAPS(cleavable amplified polymorphic sequences)标记对滇I型、野败型、红莲型及BT型恢复系基因型分析,发现这四种不同胞质的恢复系在Rf-1位点具有相同的带型,滇I型的两个保持系具有不同的带型.结果表明这四种不同胞质系统的恢复系均具有Rf-1基因.该结论与从恢复系选育的系谱分析的结果相一致的.     

淡绿叶色标记水稻光温敏核不育系开花习性

以4个淡绿叶粳稻光温敏核不育系和正常叶色粳型不育系浙农大11S(ZAU11S)及籼型不育系培矮64S(PA64S)共6个不育系为材料，在杭州自然条件下观察了上述6个光温敏核不育系的开花习性。结果表明：(1)淡绿叶色不育系同对照正常叶色不育系的叶色差异显著，易于肉眼分离；(2)淡绿叶色不育系的开花天数、开花率、柱头外露率都低于正常叶色不育系，日开花率、颖花张开时间和张开角度易受环境条件影响，表现较大差异；(3)除TS1外，其他三个淡绿叶色不育系在不育期和可育期的开花高峰都位于午前9：00或10：00，表现较强的早花特性。