發(fā)布時(shí)間：2018-05-26 02:39

  本文選題：水稻 + 株高��； 參考：《華中農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：高產(chǎn)是水稻遺傳育種的重要目標(biāo)。水稻產(chǎn)量由分蘗數(shù)、千粒重和每穗實(shí)粒數(shù)構(gòu)成,此外,株型對(duì)水稻產(chǎn)量也有重要的影響。QTL定位以及圖位克隆是發(fā)掘新基因的重要手段。利用前期02428和Teqing(TQ)的重組自交系(Recombinant Inbred Lines,RILs)群體定位到的主效株高QTL q Ph3,構(gòu)建了以TQ為供體02428背景的近等基因系,在此基礎(chǔ)上進(jìn)行q Ph3的圖位克隆以及功能研究。同時(shí),在珍汕97(Zhenshan 97,ZS97)和明恢63(Minghui 63,MH63)的RILs群體中發(fā)現(xiàn)一個(gè)株高達(dá)到160 cm的家系RI92,通過與ZS97回交的方法構(gòu)建近等基因系,并對(duì)控制株高的基因進(jìn)行圖位克隆。主要結(jié)果如下:1.對(duì)qPh3進(jìn)行遺傳分析發(fā)現(xiàn),02428等位基因型具有6.2 cm的加性效應(yīng)和3.5 cm的顯性效應(yīng),并有增加產(chǎn)量的效應(yīng)。q Ph3初步定位在WPh1到RM7389之間,通過篩選大群體中的重組單株,進(jìn)行后代測驗(yàn)確定q Ph3的基因型,把q Ph3精細(xì)定位在19-kb的區(qū)域內(nèi),兩端標(biāo)記分別為WPh10-1和RM7389,其中包含Os GA20Ox1在內(nèi)的兩個(gè)基因,親本間比較測序發(fā)現(xiàn)Os GA20Ox1的編碼區(qū)有一個(gè)SNP改變蛋白質(zhì)的編碼序列,由02428等位基因型的丙氨酸(Ala)改變?yōu)門Q等位基因型的纈氨酸(Val)。對(duì)Os GA20Ox1在近等基因系間的表達(dá)量分析發(fā)現(xiàn),02428基因型具有比TQ基因型更高的表達(dá)。2.超表達(dá)Os GA20Ox1的中花11株高增加,證明Os GA20Ox1就是qPh3中控制株高的基因。因?yàn)?Os GA20Ox1編碼GA20氧化酶,催化GA12和GA53到GA9和GA20過程中一系列的反應(yīng),所以測定了近等基因系和轉(zhuǎn)基因植株中內(nèi)源GAs的含量,結(jié)果表明,Os GA20Ox1的底物GA12和GA53在TQ近等基因系和轉(zhuǎn)基因陰性植株中得到積累,而產(chǎn)物GA20在02428近等基因系和轉(zhuǎn)基因陽性植株中明顯提高,最終導(dǎo)致了02428近等基因系和轉(zhuǎn)基因陽性植株中活性GAs不同程度地提高,印證了Os GA20Ox1是通過GAs含量影響水稻株高。3.超表達(dá)Os GA20Ox1并不增加產(chǎn)量的現(xiàn)象與近等基因系中02428等位基因型增加產(chǎn)量不一致,所以此基因組區(qū)段很可能還有一個(gè)控制產(chǎn)量的基因q Yd3,并通過挑選Os GA20Ox1純合為02428等位基因型,其他區(qū)段為分離的家系與02428回交獲得BC5F2群體用于產(chǎn)量QTL的定位和克隆。對(duì)BC5F2中q Yd3的定位分析發(fā)現(xiàn),其具有較高的增產(chǎn)潛力,q Yd3對(duì)每穗實(shí)粒數(shù)具有24粒的加性效應(yīng)和14.9粒的顯性效應(yīng),對(duì)粒寬具有0.08 mm的加性效應(yīng)和0.03 mm的顯性效應(yīng),對(duì)千粒重具有0.7 g的加性效應(yīng)和1.0 g的顯性效應(yīng),對(duì)產(chǎn)量具有4.1 g的加性效應(yīng)和4.3 g的顯性效應(yīng)。而且定位到這些的增效等位基因均是來源于02428。所以連鎖的q Yd3和q Ph3是一對(duì)相引相基因,在育種過程中只要針對(duì)一種性狀進(jìn)行選擇就可以實(shí)現(xiàn)產(chǎn)量和株高的改良。4.對(duì)RI92家系中控制株高的基因定位分析發(fā)現(xiàn),SD1很有可能是候選基因,兩親本以及RI92中SD1測序確定了RI92中在+1721到+2575之間發(fā)生了一次基因內(nèi)的重組,導(dǎo)致由ZS97的提前終止密碼子TAG改變?yōu)镸H63的TAC,從而使SD1可以翻譯成完整的GA20氧化酶蛋白,苗期多效唑的處理進(jìn)一步證明了SD1/SD1近等基因系具有更強(qiáng)的GAs合成能力。近等基因系產(chǎn)量性狀的考察顯示SD1/SD1具有增加單株產(chǎn)量的作用,但是降低收獲指數(shù)和單位面積植株數(shù)目,同時(shí)具有容易倒伏的風(fēng)險(xiǎn)。5.qPh3和SD1均在染色體端粒附近,在定位過程中,二基因所在染色體區(qū)段的重組率比基因組平均增高4倍左右,顯示重組熱點(diǎn)特征。這與染色體端粒附近一般具有重組熱點(diǎn)的觀點(diǎn)一致。
[Abstract]:High yield is an important target of rice genetic breeding. The yield of rice is composed of tiller number, thousand grain weight and the number of grain per panicle. In addition, the plant type has an important influence on the rice yield..QTL location and map cloning are the important means to excavate new genes. The recombinant inbred line (Recombinant Inbred Lines, RILs) population of early 02428 and Teqing (TQ) is used. QTL Q Ph3 of the main effect plant was located, and the near isogenic line with TQ as the donor 02428 background was constructed. On this basis, the mapping and functional study of Q Ph3 were carried out. At the same time, a family of 160 cm was found in the RILs population of Jen Shan 97 (Zhenshan 97, ZS97) and Minghui 63 (Minghui 63, MH63). The near isogenic lines were constructed and the genes of the control plant height were cloned. The main results were as follows: 1. the genetic analysis of qPh3 showed that the 02428 allele type had 6.2 cm additive effect and 3.5 cm dominant effect, and the effect of increasing yield was initially located between WPh1 and RM7389, by screening the recombinant of large population. The genotypes of Q Ph3 were determined by progeny test, and Q Ph3 was finely located in the region of 19-kb. The two ends were labeled WPh10-1 and RM7389 respectively, including two genes including Os GA20Ox1, and the encoding region of Os GA20Ox1 was found to have a coding sequence of the SNP change protein, and the 02428 allele type alanine. A) changed into the TQ allele type valine (Val). The analysis of the expression of Os GA20Ox1 in the near isogenic lines showed that the 02428 genotype was higher than that of the TQ genotype, which expressed the high expression of.2. overexpression Os GA20Ox1, which proved that Os GA20Ox1 was the gene of the control plant in qPh3. A series of reactions in the process of GA53 to GA9 and GA20, so the content of endogenous GAs in the near isogenic lines and transgenic plants was measured. The results showed that the substrate GA12 and GA53 of Os GA20Ox1 were accumulated in the TQ near isogenic lines and transgenic negative plants, while the product GA20 was obviously improved in the 02428 near isogenic lines and the transgenic positive plants. In the end, the active GAs in 02428 near isogenic lines and transgenic positive plants increased in varying degrees. It was confirmed that Os GA20Ox1 was not consistent with the increase of the yield of the high.3. overexpression Os GA20Ox1 in rice plant by GAs content, which was not consistent with the increase of the yield of the 02428 allelic genotypes in the near isogenic lines, so the genome section of this genome is likely to be still available. A gene Q Yd3, which controls yield, was selected by selecting Os GA20Ox1 homozygous as 02428 alleles. The other segments were separated from family and 02428 backcross to obtain BC5F2 population for location and cloning of yield QTL. The localization analysis of Q Yd3 in BC5F2 showed that it had higher yield potential and Q Yd3 had 24 additive effects on the number of grains per panicle. The additive effect and the dominant effect of 0.08 mm to the grain width and the dominant effect of 0.03 mm on the grain width, the additive effect and the dominant effect of 1 g on the 1000 grain weight, with the additive effect of 4.1 g and the dominant effect of 4.3 G on the yield, and the alleles of these synergies are derived from the Q Yd3 and the chain of Q Yd3 and the linkage of these genes. Q Ph3 is a pair of phase induced gene. In the breeding process, the gene localization analysis of improved.4. for yield and plant height in RI92 family can be achieved by selecting a kind of character. It is possible that SD1 is a candidate gene. Two parents and SD1 sequencing in RI92 have confirmed that RI92 has occurred between +1721 and +2575 in RI92. The recombination in the gene resulted in the change of the early terminated codon TAG of ZS97 to the TAC of MH63, so that SD1 could be translated into a complete GA20 oxidase protein. The treatment of the seedling stage of Paclobutrazol further demonstrated that the near isogenic lines of the SD1/SD1 had a stronger GAs synthesis ability. The investigation of the yield traits of the near isogenic lines showed that SD1/SD1 had an increase in single plant. The effect of yield, but reducing the harvest index and the number of plants per unit area, at the same time having the risk of easy lodging,.5.qPh3 and SD1 are all near the telomere of chromosomes. During the localization process, the recombination rate of the chromosome section of the two gene is 4 times higher than the average of the genome, which shows the feature of the recombination hot spot. This is common with the telomere of chromosomes. There is a consistent view of reorganizing hot spots.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S511

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