水稻花粉植株遗传性的观察

864丛水稻花粉植株在未经人工加倍的情况下,多倍体占1.9—8.0％,二倍体占43.3—72.0％,单倍体占20.0—48.4％。在二倍体H_1代中,可育丛为73.2—74.4％,不育丛为26.8—25.6％,其中三系杂交种的花粉植株不育丛的比例最大;H_1代育性水平不高者,在H_2代都有不同程度的恢复。杂种材料的H_2代各丛(株)系间主要经济性状都表现出多样性,并有超亲性状出现。育成品种的花粉植株多数与原品种相同或相近,只有少数材料发生明显变异。花粉植株各丛(株)系内部都表现整齐一致,没有观察到分离现象。同一块愈伤组织分化出的同丛花粉植株,多数表现相同,有少数分出两种或两种以上的不同倍性植株的混合丛。     

光敏感雄性不育水稻的体细胞与花药培养及再生植株性状表现研究

对8个光敏感雄性不育籼稻杂种一代及其亲本花药培养的研究发现,光敏不育粳稻虽然为雄性不育,但其愈伤组织诱导率与植株再生能力不在一般正常粳稻品种之下,为籼稻“新会粘”的28倍。绝大多数光敏不育之籼粳杂种的花药培养能力也是十分高的。在起源于体细胞及花药的光敏不育水稻之再生植株共1146株试管苗中,发现少数在5月下旬及6月上旬抽穗的已表现为完全雄性不育。这些材料之幼穗的第二枝梗分化时(4月下旬至5月上旬)之日长尚不足13小时,远在农垦58S的临界日长14小时之下。在8个组合的籼粳光敏杂种一代花粉植株中,有的组合完全表现为粳型(组合4及7,表2),有的完全表现为籼型(组合2及6),多数则两者兼有。在全部80个愈伤无性系中,单倍体20个,二倍体58个,多倍体2个。58个二倍体无性系申不育的为20个,占34.5%。其中在晚造短日照条件下能转变为可育的有11个,占二倍体的19%,占二倍体不育的55%。所有光敏不育花粉植株,在早造条件下I-KI之花粉染色率与结实率均为0%,在晚造(9月份之后)条件下,能转换为可育。但不同组合、不同无性系之间育性转换的频率与稳定性差异颇大。     

提高光敏感雄性不育水稻花粉单倍体育种效率及不育花粉植株育性转换特点的研

比较了（光敏 s/正常品种）F1及F2为供体亲本,对在花药培养时所获得的花粉植株中不育个体／全部花粉植株之比例的影响,结果表明,以F1为供体亲本,在所获得的二倍体花粉植株（A1）中,不育株（长日下）约占20％左右;而从F2分离的不育株为供体亲本,相应的比例为90％左右,对获得不育的花粉植株而言,供体亲本经过F2的选择,在花粉一代中可以提高育种效率3－4倍,指出,以培育光敏感雄性不育系为目的的花药培养,与一般育种之花药培养采用杂种F1为供体亲本不同,不仅应对杂种F2代在长日照条件下不育株的选择,而且应在短日照下对这种不育株作育性转换的双重选择,以这种个体作为花药培养的供体亲本,可以大大提高育种效率。在长日照下表现不育的花粉植株的育性转换具仍多样性,来自同一组合的不育花粉植株在晚造（短日照）条件下,其花粉有的染色,频率高且稳定;有的虽然可变 染色,但频率不高或不稳定或二者兼有;有些却一直不I－KI染色,或即使染色频率也在10％以下,这一结果与收集全国各地15个光敏核不育系在本所同期种植条件下的反应十分吻合,这说明通过花药培养,从特定的组合培育出所需要的光敏／光温互作或温敏型的核不育系的可能性是存在的。     

具野稻细胞质的胞质型雄性不育系的体细胞克隆变异 I.IR66707A及IR69700A体细胞克隆变异的类型

水稻雄性不育系IR66707A及IR69700A来自国际水稻所（IRRI）,它们的不育性只能保持而不能被恢复,属于胞质型雄性不育。以这两个不育系以及它们与正常品种的杂种F1为供体,经离体培养,区获得了136个体细胞克隆。在R1代中发现了三个类型的突变或变异,即：可育突变、雄性不和雌雄性均不育的变异。这一类型的可育突变有两例,其R2的可育与不可育（出现分离）之比为3：1,证实为显性单基因突变。第二类型的雄性不育又可分为两种：第一种是不育性可被恢复的,与供体不能系完全不同,其中一部分克隆的不育性不能被保持但可因环境的改变而转换为可育,另一部分克隆的不能性不受环境因素的影响（不具转换能力）,但其不育性既可恢复又可不完全保持（即F1的育性尚有分离）。第二种是不育性不能被恢复,与供体相似。育性不具转换能力的突变可望培育成为核质互作型雄性不育系,而育性可转换的突变则可望培育成为两系杂交稻中的两用核不育系。     

温光型核雄不育小麦育性转换的温度敏感期和临界温度研究

多年人工气候箱和田间分期播种试验表明,小麦温光型隐性核雄不育两用系C49S育性温度敏感期较长,从花粉母细胞形成期到成熟花粉期都有一定程度的敏感性,其中最敏感的时期有两个,即花粉母细胞减数分裂期和小孢子中期,当减数分裂期温度与小孢子中期温度均较低时,C49S表现为高度不育,当其温度均较高时,C49S表现为高度可育;C49S高度不育的临界温度是关数分裂其平均最低温度（Tmin1）、小孢子中期平均温度（T2）和平均最低温度（Tmin2）,分别为8.5℃、13.5℃和10.5℃;C49S高度可育的临界温度是Tmin1 11.5℃、T2 15.0℃、Tmin2 12.5℃;对温光型核雄不育两用系在小麦杂种优势利用中的应用价值、条件及风险进行了综合评价。     

水稻雄性不育新质源的研究──新型水稻细胞质雄性不育系马协A与野败型珍汕97A不育性的遗传学比较

采用套袋自交结实率和自然结实率为主,花粉育性和田间目测整株育性为辅的综合性状,判定新型细胞质雄性不育系马协Ａ以及它与明恢６３的杂种Ｆ_１、Ｆ_２和ＢＦ_１的植株育性,并以野败型珍汕９７Ａ作对照,比较研究了其不育性的遗传规律。结果表明,马协Ａ与珍汕９７Ａ不育性的遗传均由两对基因控制,但新型细胞质雄性不育系马协Ａ两对基因的作用方式与珍汕９７Ａ不同。前者Ｆ_２群体的育性分离符合９：３：３：１的比例,ＢＦ_１符合１：２：１的比例;后者相应群体则符合１２：３：１和２：１：１的比例,两对基因间表现为显性上位。斯米尔诺夫检验也表明马协Ａ／明恢６３和珍汕９７Ａ／明恢６３的Ｆ_２群体的结实率频率分布差异显著（Ｐ＜０．０１）。并讨论了细胞质雄性不育的遗传机理及分子基础。     

水稻体细胞无性系R_1、R_2代中的雄性育性变异观察

通过水稻幼穗培养,１９９１－１９９２两年间,在５个品种（珍汕９７Ｂ、红源Ａ、包源Ａ、Ｗ６１５４ｓ,和南广占）中共获得了５０株雄性不育变异株,其中Ｒ_１代有４８株,Ｒ_２代有２株。在Ｒ１代,共获得５２６８株再生植株,雄性不育变异的平均频率为０．９１％（０．８３－１．０８％）;在Ｒ_２代（珍汕９７Ｂ）发生雄性不育变异的频率为２％。本文报道了多种花粉败育类型之间可以相互转变现象,此外不育和可育之间亦可以相互转变。对离体培养产生的雄性不育变异株用一批现有ＣＭＳ（Ｃｙｔｏｐｌａｓｍｉｃｍａｌｅｓｔｅｒｉｌｅ）不育系的典型保持系、恢复系进行测交,结果表明,Ｗ６１５４ｓ产生的雄性育性变异株仍保持核不育的特性;红源Ａ产生的雄性育性变异株有的可能是嵌合体,有的其败育花粉类型虽发生了变化,但其恢保关系并没有改变,有的则可能已转成类似ＷＡ型的不育材料;南广占产生的典败变异株,其恢保关系类似ＷＡ型,可能属核不育转成ＣＭＳ的首例发现。     

短光低温不育水稻不育性的遗传观察

用若干常规品种与短光低温不育水稻宜ＤＩＳ配组,考察双亲及Ｆ１在自然低温影响前后的花粉育性,并于不育系稳定不育期间调查Ｆ２及Ｆ３的育性分离。结果表明：（１）低温是影响双亲及Ｆ１育性稳定性的重要生态因子;（２）Ｆ１的育性稳定性除与父本的育性稳定性有关外,可能尚与其他因素有关,这种育性稳定性表现是由可遗传因子决定的;（３）短光低温不育特性基本上符合２对独立主基因控制的遗传模式,但可能尚受到众多微效基因的     

水稻雄性不育新质源的研究──新型水稻细胞质雄性不育系马协A与野败型珍汕97A不育性的遗传学比较

采用套袋自交结实率和自然结实率为主,花粉育性和田间目测整株育性为辅的综合性状,判定新型细胞质雄性不育系马协Ａ以及它与明恢６３的杂种Ｆ１,Ｆ２和ＢＦ１的植株育性,并以野败型珍汕９７Ａ作对照,比较研究了其有性的遗传规律,结果表明,马协Ａ与珍汕９７Ａ不育性的遗传均由两对基因控制,但新型细胞质雄性不育系马协Ａ两对基因的作用方式与珍汕９７Ａ不同,前者Ｆ２群体的育性分离符合９：３：３：１的比例,ＢＦ１符合１：     

湖北光敏感核不育水稻不育性的遗传规律

本文选用5个常规粳稻品种和3小光敏感核不育系与湖北光敏感核不育水稻农垦58S杂交,用花粉育性、套袋自交结实率、自然结实率和大田目测4种育性指标同时鉴定同一植株育性,观察分析杂种后代在长日下植株育性的表现。结果表明：光敏核不育水稻不育性的遗传行为表现为受1对隐性主基因控制,不同的遗传背景对不育性的遗传行为表现有不同程度的影响。     

两个籼型水稻核不育系育性温光反应的研究

在杭州男单通过分期播种,比较了两个籼稻光温敏核不育系的育性及其转换特性。结果表明,光照长度对浙大247S和培矮64S两不育系育性表达的影响小,温度起主导作用,均属温敏型不育系,且日最低温度对不育系育性效应显著高于日平均温度和日最高温度。不育系浙大247S和培矮64S的温度敏感期分析是抽穗前3-18和6-21d,育性转换的临界日期为9月19日和9月25日,转换临界温度为25.28和25.66℃,与培矮64S相比,浙大247S不育期败育较彻底,可育期较长且自交结实率高,在杭州田间可以繁种。     

Fertile revertants from S-type male-sterile maize grown in vitro

Summary Plants were regenerated from callus cultures of maize inbred W182BN with the S(USDA) type of cytoplasmic male sterility (cms). Some regenerates from 16 of 18 separate cultures had fertile tassels. Many other regenerates, whose fertility could not be scored accurately because of abnormal plant morphology, produced fertile progeny after pollination with N cytoplasm W182BN. Revertant plants and/or progeny were obtained from all 18 cultures, which included the CA, D, LBN, and S sources of cmsS. More revertants were recovered from cultures maintained as callus for 12 months than from 3–4 month old cultures. Several types of evidence (absence of segregation for fertility after selfing or pollination of revertants with standard W182BN, pollen viability counts, failure of revertants to restore sterile cmsS lines to fertility, mitochondrial DNA analyses) indicated that the reversion to fertility involved cytoplasmic rather than nuclear alterations. All revertants examined lacked the S1 and S2 plasmid-like DNAs characteristic of the mitochondrial genome of sterile cmsS lines. Most callus cultures lost S1 and S2 after 13–20 months in vitro. No revertants were seen among thousands of W182BN cmsS plants grown from seed in the field or among plants from tissue cultures of W182BN with the C or T types of cms. The cytoplasmic revertants recovered from culture may be useful for the molecular analysis of cmsS.     

Fertility of Triticum aestivum plants derived from anther culture

The fertility of plants from wheat anther culture was studied. It was found that one half of the 36 plants with diploid root tips didn't set seeds at all, and that 41 of the 42 plants with haploid root tips were completely sterile. It was surprising that sterility was so widely distributed even among the plants with diploid root tips.     

上位性对光敏核不育水稻不育性不稳定性的影响

In this study the authors selected two indica photoperiod-sensitive genic male sterile (PSGMS) rice (Oryza sativa L.), Peiai 64S (the sterility is stable) and 8902S (the sterility is instable), and their F1,F2 populations. The genetic basis for sterile stability of PSGMS lines was studied under long-day low-temperature environments and different ecological conditions, and the genes which affected the sterile stability were mapped using RFLP analysis. The major results are as follows: The sterility instability of PSGMS lines was controlled by minor effective genes. The linkage map of Peiai 64S and 8902S has been constructed. According to Mapmaker/QTL program, seven QTLs were identified that influence the sterile stability for PSGMS lines under the long-day condition, such as L2, L3a, L3b, L5, L6, L7 and L10. They were located on the chromosomes 2, 3, 5, 6, 7, and 10 of the rice linkage map, respectively. The interaction was detected under different long-day conditions and affected the sterility stability of PSGMS. The major interactions were between additive and additive and between additive and dominance. The variances explained of epistasis were between 2.04% and 11.94%. The repeatability of the interaction was very high under different long-day conditions. The implications of these findings in hybrid rice development are also discussed.     

温敏雄性不育水稻培矮64S花药发育过程中钙的变化

采用焦锑酸钾沉淀法研究了温敏雄性不育水稻(Oryza sativa L.)培矮64S在高温引起雄性不育与正常可育花药发育过种中Ca2+的分布变化.结果表明,当培矮64S生长在较高温度条件下引起雄性不育,与可育花药相比,不育花粉母细胞中有较多的液泡、较多的Ca2+沉积和较少的线粒体,并且有较多的Ca2+沉积在不育花药的中间层、表皮层和绒毡层中.到四分体与单细胞花粉时期,不育花药的木质部细胞的次生加厚壁上有较多的Ca2+沉淀,连接组织中的Ca2+沉淀也大大增加,所有不育花粉外壁较厚而发育都不正常.在单核细胞早期,不育花粉的四分体细胞中有较明显的大液泡出现.不育花药中的Ca2+在花药发育的各时期均比可育花药要多.这些结果说明在高温生长条件下,花粉母细胞发育的异常、花药中Ca2+沉积的增加、绒毡层与花粉外壁发育的异常可能与培矮64S花粉败育相关.     

利用分子标记定位水稻野败型核质互作雄性不育恢复基因

以籼稻恢复系圭６３０与粳型广亲和品种０２４２８的Ｆ１代花药培养,获得８１个双单倍体（ＤＨ）,构建了有２３３个ＲＦＬＰ标记的分子图谱。用籼稻野败型不育系珍汕９７Ａ测定各ＤＨ系的恢复性,并将恢复性作为数量性状进行ＱＴＬ的区间作图分析,鉴别出８个基因座位,其中有２个基因座位,Ｒｆｉ－３和尾Ｒｆｉ－４,单个ＱＴＬ的基因贡献值分别是４９．６％和３５．４％,对育性恢复起主要作用,定为主效基因座位,位于第三和四染色体上,其它６个基因座位对育性恢复亦有一定的影响。表明野败型雄性不育恢复性是受主效基因和微效基因共同控制的性状。     

Comparative proteomics analysis reveals the mechanism of fertility alternation of thermosensitive genic male sterile rice lines under low temperature inducement

Thermosensitive genic male sterile (TGMS) rice line has made great economical contributions in rice production. However, the fertility of TGMS rice line during hybrid seed production is frequently influenced by low temperature, thus leading to its fertility/sterility alteration and hybrid seed production failure. To understand the mechanism of fertility alternation under low temperature inducement, the extracted proteins from young panicles of two TGMS rice lines at the fertility alternation sensitivity stage were analyzed by 2DE. Eighty‐three protein spots were found to be significantly changed in abundance, and identified by MALDI‐TOF‐TOF MS. The identified proteins were involved in 16 metabolic pathways and cellular processes. The young panicles of TGMS rice line Zhu 1S possessed the lower ROS‐scavenging, indole‐3‐acetic acid level, soluble protein, and sugar contents as well as the faster anther wall disintegration than those of TGMS rice line Zhun S. All these major differences might result in that the former is more stable in fertility than the latter. Based on the majority of the 83 identified proteins, together with microstructural, physiological, and biochemical results, a possible fertile alteration mechanism in the young panicles of TGMS rice line under low temperature inducement was proposed. Such a result will help us in breeding TGMS rice lines and production of hybrid seed.     

Ploidy stability of maize anther culture-derived callus lines

Summary Ploidy levels of 26Zea mays L. anther culture-derived callus lines of the F1 hybrids (H99 × Pa91, Pa91 × FR16, and H99 × FR16) were determined at various times after culture initiation using flow cytometry (for 21 lines) or chromosome counting of callus cells or regenerated plants (for the remaining 5 lines). Twenty of the lines remained haploid, whereas 6 were diploid. The results from flow cytometry, after examining the DNA content of 5000 nuclei of each callus line, show that each callus line consisted of homogenous haploid or diploid cells. Thus for diploid callus lines, spontaneous chromosome doubling must have occurred before or in the early stages of androgenesis, before the initiation of callus cultures. These long-term callus cultures (growing for up to 38 mo.) have stably maintained their ploidy levels so it is unlikely that the culture conditions have caused chromosome doubling. The restriction fragment length polymorphism pattern obtained with 52 to 58 markers for each diploid callus line shows that all the diploid lines are homozygous diploid so each originated from a microspore and not from diploid maternal F1 hybrid tissue.     

籼型光敏核不育水稻育性可转换性的基因定位和基因互作研究

本研究以来源于农垦58S的灿型光敏核不育系培矮64S（短日条件下育性难转换）和8902S（短日条件下育性蝗转换）及其F1,F2群体为材料,通过短日不同光温和不同生态条件4种处理,利用RFLP分子标记研究了影响光敏偿育水稻在短日条件下的育性可转换性的遗传,基因定位和基因互作,主要结果表明：影响光敏不育水稻的育性可转换性表现为微效基因的作用,定位了7个控制光敏核不育水稻的育性可转换性QTL,即S2,S3a,S3b,S5,S8和S10,揭示了基因互作真实存在于光敏核不育水稻中,基因互作形式和互作类型对光敏核不育水稻的育性可转换性的影响表现多种多样,不同类型的基因互作所解释的遗传变异处于2．15％－10．07％之间。     

DNA methylation changes in photoperiod-thermo-sensitive male sterile rice PA64S under two different conditions

Epigenetic modification can occur at a high frequency in crop plants and might generate phenotypic variation without changes in DNA sequences. DNA methylation is an important epigenetic modification that may contribute to environmentally-induced phenotypic variations by regulating gene expression. Rice Photoperiod-Thermo-Sensitive Genic Male Sterile (PTGMS) lines can transform from sterility to fertility under lower temperatures and short-day (SD) conditions during anther development. So far, little is known about the DNA methylation variation of PTGMS throughout the genome in rice. In this study, we investigated DNA cytosine methylation alterations in the young panicles of PTGMS line PA64S under two different conditions using methylation sensitive amplified polymorphism (MSAP) method. Compared with the DNA methylation level of PA64S under lower temperatures and SD conditions (fertility), higher methylation was observed in PA64S (sterility). The sequences of 25 differentially amplified fragments were successfully obtained and annotated. Three methylated fragments, which are homologous to D2, NAD7 and psaA, were confirmed by bisulfite sequencing and their expression levels were also evaluated by qPCR. Real time quantitative PCR analysis revealed that five of the six selected methylated genes were downregulated in PA64S (sterility). These results suggested that DNA methylation may be involved in the sterility–fertility transition of PA64S under two different environmental conditions.