【作者】 罗红兵；

【导师】 黄璜；

【作者基本信息】 湖南农业大学 ， 作物栽培学与耕作学， 2002， 博士

【摘要】 通过杂种后代的自交分离，选育成新的细胞质雄性不育系GDS及其相应保持系BG。本研究对GDS细胞质雄性不育系的基本特性、胞质分类、细胞学特征、抗病特性、可恢复性及恢复性的遗传性和稳定性、GDS不育系不同类型恢复系的RAPD分析等进行了研究，同时研究了不同类型的细胞质雄性不育系对不同来源的恢复系的反应，以便为GDS细胞质雄性不育系的利用提供可靠的证据，并为玉米细胞质雄性不育系恢复系的选育提供新途径和理论依据。研究结果如下： 1．玉米GDS细胞质雄性不育系的基本特征 1．1 不育性状的稳定性：不育系GDS、GDS（478）不育性十分稳定。环境中的温度、光照不能影响其育性表现。 1．2 形态特征：GDS及GDS（478）在不同生态条件下，雄穗花颖都关闭，不开裂。GDS花药长度为其保持系花药长度的58％。极少有花粉，花粉为碘败型花粉雄性不育类型。 1．3 穗部性状特性：不育系GDS、GDS（478）和其相应保持系BG、478在同一生态条件下，在雄穗主轴长度、主穗小穗数、分枝数、总小穗数等雄穗性状方面，同一不育系与其相应保持系差异不大，但在不同生态条件下，各性状差异较大，在三亚冬播，雄穗分枝数比长沙春、秋播时少。 1．4 光合速率特性：不育系GDS、GDS（478）在大喇叭期、抽雄吐丝期、灌浆期三个时期的光合速率均比其相应保持系BG、478强。三个时期中，以抽雄吐丝期光合速率最强，此期GDS（478）、478、GDS、BG的光合速率值分别为33.91、33.85、32.22、30.26μmol·m^-2·s^-1。 2．GDS细胞质雄性不育系分类 2．1 雄性不育系的田间育性表现：待测不育系GDS、GDS（478）与11个对照不育系除对照系中S型不育系C836D、C836H在三亚、长沙育性不稳定，待测不育系和对照中5个T型不育系、2个C型不育系和其他2个S型不育系在两地不育性状稳定。 2．2 GDS胞质恢复专效性测定：恢复系C936D（基因型Rf₁Rf₁Rf₂Rf₂ rf₃rf₃ RfC眈）、C936F（基因型巾rfl听叫r方巾肌m）能使GDS、GDS（478）育性恢复， 而恢复系C736CB p助叫阶)、C936Km人队叫叫)不自恢复GDS、GDS（478） 的育性，由此可以推断GDS、GDS仟78)属于C型细胞质雄性不育系。 23 GDS胞质不育系PCR快速归类：用3组特异性5！物对待测不育系GDS、 GDS（478和己知不育胞质类型B37－Veins－乃、B37－C（cm－q、B37－s（cm－S）及可 育系B37－N进行PCR扩增，结果GDS、GDS以78）所扩增出的产物与B37－C（cm－C） 产生的带形分子量大小相同，表明GDS、GDS以78）M于C型不育胞质，分类结巴—————一、 果与恢复专效性测定结果相吻合。 3．GDS胞质雄性不育的细胞学特征和抗病性鉴定 3．IGDS胞质雄性不育系小抱子败育特征：GDS不育系小抱子败育发生的时间 较早：在花粉母细胞减数分裂前期1就表现出败育的特征，不能通过减数分裂。 败育呈两种形式，第一种类型绒毡层异常膨大，花粉母细胞浓缩解体。第二种类 型是绒毡层液泡化程度低，花粉母细胞早期开始浓缩解体。这一类型未见报道。 32 GDS不育胞质对大小斑病的抗性：人工接种大、小斑病病菌，结果表明GDS 不育系对大斑病是高抗的。而对小斑病病菌反应与保持系BG一致。 4．GDS细胞质雄性不育系恢复性研究结果＠4工 天然杂交法选择GDS细胞质雄性不育系的恢复系：从M不育天然杂交株 后代可育株中连续自交分离，筛选到GDS胞质不育系的强恢复系H53、H54、 H57、H58。 4．2 GDS不育胞质杂交种和正常胞质（保持系BG）杂交种比较：GDS不育胞质与 正常胞质组配的杂交种，其产量无明显差异，GDS胞质杂交种平均小区产量为 4．27kg，正常胞质杂交种平均小区产量4．22kg。GDS胞质与正常胞质杂交种在株 高、穗位高、播种一抽雄天数、播种一吐丝天数、穗长、穗粗、穗行数、行粒数、 千粒重等项目中，均无显著差异，而不同的恢复系（H53、H54、H58）配制出的组 合除穗粗无显著差异，穗行数差异显著，其它性状均达极显著水平。GDS X H58 在品比试验中，其平均产量 504．2kg／667m‘，比对照掖单 13号增产 4．28％，产量 ＠、·一’““—一 差异未达显著水平。 4．3 GDS不育恢复性的遗传：组合GDSXH53、GDSXH54、GDSXT01三个组 合可育株与不育株 FZ的分离比例为 3二，而其与保持系测交后代呈 1＊的分离比 例，适合性测定结果表明H53、H54、T01具有一对恢复基因。但GDS XN0lFZ 及（GDS XN01）XBG中不育株比例严重偏高。 11 4．4 玉米 GDS不育胞质杂交种雄花育性恢复稳定性：杂交组合 GDS X H58、GDS XT01、GDSXN01恢复度高更多还原

【Abstract】 A cytoplasmic male sterile (cms) line GDS in Maize(Zea mays) and its maintainer BG were selected from segregation of self-pollination. This study focused on the following points: a) characteristics of GDS in growth, cytoplasm type via a Polymerase Chain Reaction (PCR) protocol, cytological property, and disease resistance; b) restorer and its genetic background and FI fertility stability; and c) various types of GDS restorers via a Random Amplified Polymorphic DNA (RAPD). Simultaneously, responses of various cms types (cms-T, -C, and -S) to restorers from different backgrounds were investigated, which could be important to utilize cms in seed production and provided new methods and guidelines for selecting suitable restorers. The main conclusions can be summarized as follows. 1. Characteristics of male sterile line1.1 Stability of male sterilityMale sterile lines GDS and GDS(478) were stable across testing seasons and locations. This indicated that this kind of male sterility was independent of environment condition, e.g., temperature and light.1.2 Configuration aspectsAmong various conditions, tassel and anthers from cms lines GDS and GDS(478) were closed during maturity. The length of anther from cms line GDS was 58% of that from normal line (maintainer). There is no pollen in most of anthers.1.3 Tassel growthThe differences in main axis length, spikelets per main axis, branches, and total spikelets were marginal between cms lines GDS, GDS(478) and their maintainers. These parameters did varied with growing seasons. Tassel branches were much less in winter at Sanya than in spring and autumn at Changsha.1.4 Photosynthesis ratePhotosynthesis rate was monitored at three main stages, most of which were higher in cms lines GDS and GDS(478) than their maintainers. The highest photosynthesis rate was detected at silking (tasselling) period as 33.91, 33.85, 32.22, and 3Q.26umol-m~2-s~l for GDS(478), 478, GDS, and BG lines, respectively.2. Discrimination of GDS male sterile type2.1 Field observationGDS, GDS(478), and other 11 cms (5 cms-T, 2 cms-C, and 4 cms-S) lines as CK were tested for their fertile behavior. Except C836D (S) and C836H (S), all other lines were male sterile in both Sanya and Changsha over seasons.2.2 Restorers for GDS and its deduced male sterile typeBoth lines C936D (RfiRfiRf2Rf2 rf3rf3 RfCRfC) and C936F (rfirf1Rf2Rf2rf3rf3RfCRfC) could restore GDS and GDS(478), while C736CB (RfiRfiRf2Rf2) and C936K (RfiRfiRf2Rf2) could not do. It is deduced that GDS and GDS(478) were grouped into cms-C type.2.3 Discrimination of male sterile type from a rapid PCR procedureUsing 3 pairs of specific primers, GDS, GDS(478),B37-N(Normal), B37-T(cms-T), B37-C(cms-C), and B73-S(cms-S) were tested via a PCR protocol. There were similar bands from GDS and GDS(478) lines as B73-C. Thus, GDS and GDS(478) should be cms-C type, which is consistent with the test from specific restorers.3. Cytological characteristic and disease resistance3.1 Characteristics of microspore abortion in male sterile GDSMicrospore abortion happened relatively early in male sterile GDS, shown in early stages of microspore maternal cell meiosis. This means, microspore maternal cell failed to meiosis. There were two types of abortion, i.e., a) fine layers expanding abnormally and microspore maternal cells condensing to disjoint and b) fine layers vacuolifing to a very low extent and maternal cells in pollens beginning to condense and disjoint. There is no report for the second type up to now.3.2 Disease resistanceWith manual inoculation pathogen, cms GDS line was highly insensitive to Nothern blight of corn, but both cms line CDS and its maintainer BG were similar reaction to Southern blight of corn.4. Restorer for cms CDS4.1 Natural selecting restorerFrom segregation of M sterile plant which was in natural cross-pollination, the fertile plants were selected and self-pollinated continuously. Four lines, named as H53, H54, H57, and H58 were obtained for restoring GDS.4.2 Compa更多还原