【作者】 康公平；

【导师】 陈良碧；

【作者基本信息】 湖南师范大学 ， 植物学， 2012， 博士

【摘要】 水稻正反交杂种优势存在不一致现象、即细胞质效应,三系杂交水稻恢复系与保持系杂交的优势大多大于与不育系的杂交优势,即不育细胞质存在负效应。两系杂交稻不受细胞质约束,可以培育出各种不同细胞质类型的核不育系和高产组合,然而两系杂交稻选育都忽视了细胞质效应。本研究以细胞质来源于广东珠江流域的广陆矮四号、江苏长江中下游的9311、菲律宾国际水稻所的R402、日本的日本晴、南美的巴西陆稻、北美的Lemont、我国北纬18.73。的普通野生稻(野败)V20A和北纬26.50°的普通野生稻茶陵野生稻为细胞质供体,以株1S、准S为细胞核供体,经连续回交12次,构建两套1种核9种细胞质的同核异质温敏核不育系,并与父本9311、527、R819、师大21杂交制备72个同核异质组合,在相同栽培条件下比较同核异质温敏核不育系及同核异质组合生物学性状,探讨不同类型细胞质的效应以及与核、质互作关系,旨在为两系法杂交水稻选育最佳不育系及核、质互作组合提供理论指导。主要结果如下：1、对cpDNA9个片段共计9080个碱基的比对分析表明,8个细胞质供体以及细胞核供体株1S、准S的cpDNA之间都存在遗传差异,并根据碱基差异建立了各种细胞质的cpDNA分子指纹。2、利用ISSR、SRAP和TRAP三种分子标记方法对细胞核供体核DNA分析表明,细胞核供体株1S与准S的核DNA存在遗传差异,并建立了细胞核供体核DNA分子指纹。3、同核异质不育系的制备以及代换系纯度的分子鉴定构建了两套1种核9种细胞质的同核异质温敏核不育系材料,经516对SSR引物标记对同核异质不育系细胞核进行检测,结果表明所转育的不育系材料与细胞核供体完全一致,且同类不育材料在大田的农艺性状完全一致,前后代之间没有性状分离。4、细胞质对不育系及杂交组合主要农艺性状的影响细胞质对不育系或组合的株叶型、生育期等都有不同程度影响。例如C5(为了保密,细胞质源名称省略,下同)细胞质能显著增加不育系的剑叶面积、宽度：以C5为细胞质源制备株1S、准S同核异不育系的剑叶面积比对照(株1S、准S)大2.64cm2、4.77cm2；剑叶宽度比对照宽0.19cm、0.15cm。 C7细胞质显著降低组合的株高：具C7细胞质源的组合株高比对照矮0.4～11.Ocm。具C7细胞质源的组合全生育期比对照缩短2.2～5.2天,具C3细胞质源的组合全生育期比对照长0.4～3.1天。5、细胞质对不育系及杂交组合主要经济性状的影响细胞质对不育系或组合的有效穗、穗长、颖花数(穗粒数)、结实率等都有不同程度影响。C7细胞质能降低不育系、组合的穗长和颖花数,以C3为细胞质的组合结实率较其他细胞质源组合低2.0-8.1个百分点。C4细胞质对部分组合有效穗有正效应。6、细胞质对不育系光合特性影响在株1S核源中,以C5为细胞质源的不育系净光合速率、Fv’/Fm’、 PhiPS2、qP显著大于对照,而以C3为细胞质源的不育系则显著小于对照,在准S核源中,以C8为细胞质源的不育系净光合速率、Fv’/Fm’ PhiPS2、qP显著大于对照,而以C7为细胞质源的不育系则显著小于对照。在株1S、准S两个核源中,换质后的不育系光合色素含量、表观量子效率、光补偿点、光饱和点、ETR与对照差异不显著。7、细胞质对杂交组合生物学优势的影响C2、C4、C5细胞质对杂交组合的地上部分干重有正效应：具C2细胞质源的晚稻组合地上部分干重高出对照7.6%、1.8%(前者为株1S组合,后者为准S组合,下同)；具C4细胞质源的早稻组合地上部分干重高出对照2.9%～7.7%、1.7%～4.0%；C5细胞质源的组合地上部分干重高出对照2.3%～16.8%。而具C3细胞质源的组合地上部分干重低于对照6.7%～18.3%、16.0%～17.6%；具C7细胞质源的早、中稻组合低于对照3.9%～9.6%、18.0%～19.0%。8、细胞质对温敏核不育系临界不育温度的影响株1S被更换不同细胞质后,花粉不育的稳定性最差的是C6细胞质配制的不育系,临界不育温度超过了23.5℃,不适宜在长江中下游稻区应用。以C1、C8为细胞质源的不育系其不育的稳定性比株1S有所降低,但可繁性有所提高。以C2、C3为细胞质的不育系其不育的稳定性比株1S更高,但可繁性更低。在更换不同细胞质的准S系列不育系中,导致临界不育温度显著升高、花粉不育的稳定性最差的是C7细胞质,而C2、C3细胞质能显著提高不育性的稳定性,使不适宜在长江中下游稻区应用的准S达到安全应用标准。综上所述,同核异质不育系及同核异质组合的生物学性状主要受核基因控制,但细胞质效应和质核互作效应对部分性状也有显著影响。C2细胞质是选育温敏核不育系最适宜的细胞质,有利于降低临界不育温度,提高杂交制种的安全性。利用C7细胞质有利于选育早熟早稻组合。C2、C4、C5为细胞质有利于选育高产组合。更多还原

【Abstract】 Researchs indicate that the heterosis of the F1progenies of reciprocal crosses exist some difference,this phenomenon is cytoplasmic effect.The heterosis of the cross combination of the restorer line with the maintainer line almost is better than that of the cross combination of the restore line with the sterile line in three-line hybrid rice,this phenomenon is the negative effect of sterile cytoplasm.Two-line hybrid rice systems are more advantageous than three-line hybrid rice systems because of the absence of any cytoplasmic restriction on breeding a nuclear sterile line and high-production combinations,howereve,the breeding of two-line hybrid rice all ignore the cytoplasmic effect.In this project,common wild rice,cultivated rices that originate from the Northen America,the Southern America,Japan,the Philippines were used as the cytoplasmic donors.Zhu-1S and ZhunS were used as the nuclear donors. The cytoplasmic donors were used as female parents in crosses with the nuclear donors, as males.The mononuclear-heteroplasmic sterile lines were prepared via15backcrosses using the original male as recurrent parent.The preliminary results indicate that cytoplasm affect the agronomic traits of the mononuclear-heteroplasmic cross combinations which were prepared by crossing the mononuclear-heteroplasmic sterile lines and four male parents.The genome of mononuclear-heteroplasmic sterile lines will be identified by the molecular markers of SSR in this Study.Base on the identity of the nuclei of mononuclear-heteroplasmic sterile lines,effects of cytoplasm on the mononuclear-heteroplasmic sterile lines and the mononuclear-heteroplasmic cross combinations will be studied,the photosynthetic characteristic of the significant difference of biomass yield of the heteroplasmic sterile and heteroplasmic cross combination will be analysed.This study aim to construct a series of special genetic materials for studing the cytoplasmic effects,and promote promote the further development of the theory of cytoplasmic effect,and provide theoretical support for the best combination of nucleu-cytoplasm in two-line hybrid rice.1、Using9cpDNA sequences have established cytoplasmic molecular fingerprint of8cytoplasmic donors and2nuclear donors, the results showed that ten kinds of cytoplasm had varying degrees of genetic difference.2、The genomes of nuclear donors were analysed using inter-simple sequence repeat(ISSR), Sequence-related amplified polymorphism(SRAP) and target region amplification polymorphism(TRAP) molecular markers. The results showed that two nuclear donors had varying degrees of genetic difference,and the molecular fingerprints of nuclear donors had established.3、The mononuclear-heteroplasmic thermo-sensitive genic male sterile (TGMS) lines were confirmed that their nucleus were identical with the donors of nucleu respectively using516pairs of SSR markers. Meanwhile, no segregation of characters was found in BC11F1-BC12F1 generations of the same substitution line in field experiments.4、The effect of C5cytoplasm on area and width of the flag were positive,whereas the effect of C8cytoplasm on area and width of the flag were negative. The plant height of cross combinations with C7cytoplasm were0.4～11.0cm lower than those of controls. The full growth period of cross combinations with C7cytoplasm were2.2～5.2d shorter than those of controls, The full growth period of cross combinations with C3cytoplasm were0.4～3.1d longer than those of controls.5、In two nuclear backgrounds, the effect of C7cytoplasm on panicle length, spikelet No./panicle of TGMS line were negative. The filled-grain of cross combinations with C3cytoplasm were2.0～8.1percentage points lower than those of controls.The effect of C4cytoplasm on panicle No./plant of cross combinations were positive.6、In two nuclear backgrounds, the differences that the content of photosynthetic pigments, AGY, light compensation point, light saturation point, and ETR of among the mononuclear-heteroplasmic lines were not significant. In ZhulS nuclear background,the net photosynthetic rate, Fv’/Fm’, PhiPS2, qP of the substitution line with C5cytoplasm are significantly higher than those of Zhu1S, whereas those of the substitution line with C3cytoplasm were significantly lower than the those of Zhu1S.In ZhunS nuclear background, the net photosynthetic rate, Fv’/Fm’, PhiPS2, qP of the substitution line with C8cytoplasm were significantly higher than those of Zhu1S, whereas those of the substitution line with C7cytoplasm were significantly lower than those of ZhulS.7、The effect of C2、C4and C5cytoplasms on the dry weights of above-ground of cross combinations were positive. The dry weights of above-ground of late cross combinations with C2cytoplasm were1.8%～7.6%higher than those of controls, the dry weights of above-ground of early cross combinations with C4cytoplasm were1.7%～7.7%higher than those of controls, the dry weights of above-ground of cross combinations with C5cytoplasm were2.3%～16.8%higher than those of controls. But the dry weights of above-ground of cross combinations with C3cytoplasm were6.7%、18.3%lower than those of controls, The dry weights of above-ground of early-season and mid-season cross combinations with C7cytoplasm were3.9%～19.0%lower than those of controls.8、In the Zhu-1S nuclear background, the least stable pollen sterility was observed in the substitution line with C6cytoplasm, and its critical sterility temperature points(CSTP) were above23.5℃. Hence, this cytoplasm were unsuitable in the ecological region in the middle and lower reaches of the Yangtze River. The substitution lines with C3and C2cytoplasms had more stable pollen sterilities than Zhu-1S, but their fecundities were lower than that of Zhu-1S. In the ZhunS nuclear background, the substitution line with C7cytoplasm had a higher CSTP and the least stable pollen sterility. The substitution lines with C2and C3cytoplasms had the most stable pollen sterility and were suitable for application in the ecological region in the middle to lower reaches of the Yangtze River.All the analysis indicated that the biological traits of mononuclear-heteroplasmic TGMS lines and mononuclear-heteroplasmic cross combinations were controlled by nucleu,but the effects of cytoplasm and interaction of cytoplasm and nucleu on some traits are significant also.In this study,the C2cytoplasm was the most suitable cytoplasm for breeding TGMS lines. The C7cytoplasm was the best cytoplasm for breeding the early season hybrid rice combinations. The C2、C4and C5cytoplasms were excellent candidates for breeding the high-yield hybrid rice combinations.更多还原