棉花零式果枝性狀候選基因功能的初步分析
發(fā)布時(shí)間：2018-08-24 12:41
【摘要】：棉花是重要的經(jīng)濟(jì)作物,其株型結(jié)構(gòu)對(duì)產(chǎn)量具有較大影響。目前生產(chǎn)上應(yīng)用的大部分陸地棉品種為無(wú)限果枝類型,果枝包含多個(gè)果節(jié),可以無(wú)限生長(zhǎng);而零式果枝突變體通常表現(xiàn)為一節(jié)有限生長(zhǎng),甚至花芽直接分化于主莖葉腋處。零式果枝突變具有早熟、株型緊湊、花期集中的特點(diǎn),因此,研究零式果枝突變對(duì)研究果枝發(fā)育,調(diào)控株型結(jié)構(gòu),使棉花株型緊湊,更加適于高密度種植和機(jī)采等具有重要的理論和實(shí)踐意義。本研究組前期利用分子標(biāo)記將海島棉零式果枝基因定位于D7染色體上約600kb的區(qū)間內(nèi),其中包含有1個(gè)金魚草CEN(CENTRORADIALIS)同源基因,是零式果枝基因可能的候選基因,命名為GhCEN。本研究對(duì)GhCEN利用了序列分析及擬南芥異源表達(dá)等手段,來(lái)驗(yàn)證GhCEN為控制零式果枝的重要基因。主要結(jié)果如下:1零式果枝性狀表現(xiàn)為隱性遺傳。通過(guò)對(duì)不同果枝類型的棉花品種的GhCEN基因的克隆、測(cè)序及序列比對(duì)發(fā)現(xiàn),零式果枝材料新海18的GhCEN在P133S位存在單堿基的突變,此位點(diǎn)為植物CEN同源基因的保守位點(diǎn),其突變可能導(dǎo)致GhCEN功能喪失,和單隱性基因的遺傳模式相一致。2定量RT-PCR分析GhCEN基因的時(shí)空模式。結(jié)果表明,GhCEN在根中的表達(dá)量較高,莖尖中也有一定程度的富集;葉片中GhCEN基因mRNA豐度極低,并不隨葉齡發(fā)生變化。不同光周期處理結(jié)果表明:長(zhǎng)日照處理下(16h),根和葉片都是在光培養(yǎng)條件下進(jìn)行GhCEN的合成,并呈現(xiàn)晝夜節(jié)律變化;短日照處理下(8h),GhCEN基因在葉片的表達(dá)量顯著高于根中表達(dá)量。3轉(zhuǎn)擬南芥功能驗(yàn)證。采用35S啟動(dòng)子在擬南芥中過(guò)表達(dá)GhCEN基因和突變基因cen都可以導(dǎo)致擬南芥的提早開(kāi)花。棉花中零式果枝材料表現(xiàn)為早花,因此GhCEN基因在擬南芥和棉花中可能受到不同的遺傳調(diào)控。
[Abstract]:Cotton is an important cash crop, its plant type structure has great influence on yield. At present, most upland cotton varieties used in production are infinite fruit branch type, the fruit branch contains more than one fruit node and can grow infinitely, while the zero-type fruit branch mutant usually shows a limited growth, even the flower bud differentiates directly in the main stem and leaf axils. The zero-type branch mutation has the characteristics of early maturity, compact plant type and concentrated flowering period. Therefore, the study of zero-type branch mutation on the study of fruit branch development, regulation of plant type structure, making cotton plant type compact, More suitable for high density planting and mechanical harvesting has important theoretical and practical significance. This study used molecular markers to locate the zero-type fruit-branch gene in the region of 600kb on chromosome D7, which contained a CEN (CENTRORADIALIS) homologous gene, which was a possible candidate gene for zero-type fruit-branch gene, named GhCEN.. In this study, sequence analysis and heterologous expression of Arabidopsis thaliana were used to identify GhCEN as an important gene to control zero-type fruit branch. The main results were as follows: 1. By cloning, sequencing and sequence alignment of GhCEN gene of cotton varieties with different fruit-branch types, it was found that the GhCEN of the zero-type fruit branch material Xinhai 18 had a single base mutation at the P133s site, which was the conserved site of the plant CEN homologous gene. The mutation may lead to the loss of GhCEN function, which is consistent with the genetic pattern of single recessive gene. 2. Quantitative RT-PCR is used to analyze the temporal and spatial pattern of GhCEN gene. The results showed that the expression of GhCEN was higher in the root and the stem tip was also enriched to some extent, and the mRNA abundance of the GhCEN gene in the leaves was very low and did not change with the leaf age. The results of different photoperiod treatments showed that under long sunlight (16h), root and leaf were synthesized under light culture condition, and the circadian rhythm was observed. (8 h) GhCEN gene expression in leaves was significantly higher than that in roots (3. 3%) and the function of Arabidopsis thaliana was verified. Overexpression of GhCEN gene and mutant gene cen in Arabidopsis thaliana by 35s promoter can lead to early flowering of Arabidopsis thaliana. The zero-type fruit-branch materials in cotton showed early flowering, so the GhCEN gene may be controlled by different genetic regulation in Arabidopsis thaliana and cotton.
【學(xué)位授予單位】：塔里木大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S562
[Abstract]:Cotton is an important cash crop, its plant type structure has great influence on yield. At present, most upland cotton varieties used in production are infinite fruit branch type, the fruit branch contains more than one fruit node and can grow infinitely, while the zero-type fruit branch mutant usually shows a limited growth, even the flower bud differentiates directly in the main stem and leaf axils. The zero-type branch mutation has the characteristics of early maturity, compact plant type and concentrated flowering period. Therefore, the study of zero-type branch mutation on the study of fruit branch development, regulation of plant type structure, making cotton plant type compact, More suitable for high density planting and mechanical harvesting has important theoretical and practical significance. This study used molecular markers to locate the zero-type fruit-branch gene in the region of 600kb on chromosome D7, which contained a CEN (CENTRORADIALIS) homologous gene, which was a possible candidate gene for zero-type fruit-branch gene, named GhCEN.. In this study, sequence analysis and heterologous expression of Arabidopsis thaliana were used to identify GhCEN as an important gene to control zero-type fruit branch. The main results were as follows: 1. By cloning, sequencing and sequence alignment of GhCEN gene of cotton varieties with different fruit-branch types, it was found that the GhCEN of the zero-type fruit branch material Xinhai 18 had a single base mutation at the P133s site, which was the conserved site of the plant CEN homologous gene. The mutation may lead to the loss of GhCEN function, which is consistent with the genetic pattern of single recessive gene. 2. Quantitative RT-PCR is used to analyze the temporal and spatial pattern of GhCEN gene. The results showed that the expression of GhCEN was higher in the root and the stem tip was also enriched to some extent, and the mRNA abundance of the GhCEN gene in the leaves was very low and did not change with the leaf age. The results of different photoperiod treatments showed that under long sunlight (16h), root and leaf were synthesized under light culture condition, and the circadian rhythm was observed. (8 h) GhCEN gene expression in leaves was significantly higher than that in roots (3. 3%) and the function of Arabidopsis thaliana was verified. Overexpression of GhCEN gene and mutant gene cen in Arabidopsis thaliana by 35s promoter can lead to early flowering of Arabidopsis thaliana. The zero-type fruit-branch materials in cotton showed early flowering, so the GhCEN gene may be controlled by different genetic regulation in Arabidopsis thaliana and cotton.
【學(xué)位授予單位】：塔里木大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S562
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