發(fā)布時(shí)間：2018-04-25 16:35

  本文選題：小桐子 + 非生物脅迫　； 參考：《昆明理工大學(xué)》2017年碩士論文

【摘要】：小桐子(Jatrophacurcas L.)屬于大戟科麻風(fēng)樹(shù)屬,是近年來(lái)受到廣泛關(guān)注的一種能源植物。為了不與糧食作物爭(zhēng)地和改善生態(tài)環(huán)境的需要,小桐子一般被種植在貧瘠、干旱的土地上,面臨多種逆境脅迫,極大地影響了它的產(chǎn)量與油質(zhì),限制了它的地域分布,故對(duì)其抗逆性的研究具有重要意義。本研究從小桐子成熟的葉片中克隆獲得5個(gè)與其抗逆相關(guān)基因的cDNA全長(zhǎng),進(jìn)行了生物信息學(xué)分析,并結(jié)合qRT-PCR方法研究了以上5個(gè)基因在小桐子不同組織、種子不同發(fā)育階段和不同非生物脅迫處理下的表達(dá)模式,為有效挖掘和利用小桐子的抗性優(yōu)良基因提供參考。本研究獲得的主要結(jié)果如下:1、克隆得到JcNF-YB3和JcNF-YB10基因的CDS序列,分別包含549bp和528bp完全閱讀框(ORF),編碼182和175個(gè)氨基酸,都與油桐NF-YB家族基因序列一致性最高且親緣關(guān)系最近。定量分析表明,JcNF-YB3和JcNF-YB10在小桐子根、莖、葉、花、果皮和種子中都有表達(dá),JcNF-YB3在根中表達(dá)量最高,JcNF-YB10在種子中表達(dá)量最高。兩個(gè)基因在種子發(fā)育期的表達(dá)量變化趨勢(shì)一致,但是JcNF-YB10在種子整個(gè)生長(zhǎng)發(fā)育期的表達(dá)量都要高于JcNF-YB3。JcNF-YB3和JcNF-YB10都在NaCl、PEG、4℃低溫、機(jī)械損傷和ABA處理后表達(dá)量不同程度上調(diào)。2、克隆得到JcGSK基因的CDS序列,包含1230bpORF,編碼409個(gè)氨基酸,與巴西橡膠樹(shù)GSK蛋白的氨基酸序列一致性最高(94%)且親緣關(guān)系最近。組織表達(dá)結(jié)果顯示,JcGSK在小桐子根、莖、葉、花、果皮和種子中都有表達(dá),在根中表達(dá)量最高,種子中的表達(dá)量次之,在種子發(fā)育過(guò)程中表達(dá)量的變化與種子生長(zhǎng)發(fā)育趨勢(shì)基本一致。小桐子幼苗在NaCl、PEG、4℃低溫、機(jī)械損傷和ABA處理后JcGSK表達(dá)量不同程度上調(diào)。3、克隆得到JcYkt6基因的CDS序列,包含600bpORF,編碼199個(gè)氨基酸,與香瓜和黃瓜的Ykt6蛋白序列一致性最高(91%)且親緣關(guān)系最近。定量分析結(jié)果顯示,JcYkt6基因在小桐子根、莖、葉、花、果皮和種子中都有表達(dá),在種子中表達(dá)量最高。在種子發(fā)育期表達(dá)量變化過(guò)程與種子中儲(chǔ)藏蛋白質(zhì)合成趨勢(shì)基本一致。NaCl、PEG、4℃低溫、機(jī)械損傷和ABA處理下JcYkt6表達(dá)水平都不同程度上調(diào)。4、克隆得到Jc5β-StR基因的CDS序列,包含1173 bp ORF,編碼390個(gè)氨基酸,與蓖麻5β-StR蛋白序列一致性最高(87%)且親緣關(guān)系最近。Jc5β-StR是一個(gè)短鏈還原酶,屬于SDR超家族。組織表達(dá)結(jié)果顯示Jc5β-StR在小桐子根、莖、葉、花、果皮和種子中都有表達(dá),在種子中表達(dá)量最高,在種子生長(zhǎng)發(fā)育期表達(dá)量變化與種子生長(zhǎng)發(fā)育的趨勢(shì)基本一致。NaCl、PEG、4℃低溫、機(jī)械損傷和ABA處理下,Jc5β-StR的表達(dá)量都不同程度上調(diào)�？傊�,JcNF-YB3、JcNF-YB10、JcGSK、JcYkt6和Jc5β-StR都具有組織表達(dá)特異性,可能參與調(diào)控小桐子器官生長(zhǎng)發(fā)育,雖然在不同非生物脅迫條件下的表達(dá)模式不同,但是它們都能夠參與小桐子對(duì)于非生物脅迫的應(yīng)答,表明它們?cè)谛⊥┳拥目鼓鏅C(jī)制上也起到一定作用。
[Abstract]:Jatrophacurcas L. It belongs to the genus Euphorbiaceae, and is a kind of energy plant which has received extensive attention in recent years. In order not to compete with food crops for land and to improve the ecological environment, jatropha is generally planted on barren, arid land and is subject to various stresses, which greatly affect its yield and oil quality and limit its geographical distribution. Therefore, the study of its resistance is of great significance. In this study, the full-length cDNA of five genes related to stress resistance were cloned from mature leaves of Paulownia chinensis. The bioinformatics analysis was carried out, and the above five genes were studied in different tissues of Paulownia cuspidata with qRT-PCR method. The expression patterns of seeds at different developmental stages and under different abiotic stress provide a reference for the effective mining and utilization of the fine genes of resistance of Paulownia chinensis. The main results of this study were as follows: 1. The CDS sequences of JcNF-YB3 and JcNF-YB10 genes were cloned, including 549bp and 528bp complete reading frame, encoding 182 and 175 amino acids respectively. Quantitative analysis showed that JcNF-YB3 and JcNF-YB10 were expressed in the roots, stems, leaves, flowers, pericarp and seeds of Paulownia cuspidata. JcNF-YB3 expressed the highest amount in the root and JcNF-YB10 was the highest in the seeds. The expression of JcNF-YB10 was higher than that of JcNF-YB3.JcNF-YB3 and JcNF-YB10 at 4 鈩，

本文編號(hào)：1802122