棉花多逆境響應(yīng)基因的挖掘和功能驗(yàn)證
發(fā)布時(shí)間：2018-11-07 06:56
【摘要】：植物通過復(fù)雜而精細(xì)的調(diào)控網(wǎng)絡(luò)感知外界的各種環(huán)境變化,激活相關(guān)信號路徑,抵御逆境脅迫,但同時(shí)往往伴隨著生長發(fā)育進(jìn)程的改變并影響產(chǎn)量。由于基因調(diào)控網(wǎng)絡(luò)的復(fù)雜性,部分基因位于多個信號路徑的交叉點(diǎn)形成樞紐基因,并在多個信號轉(zhuǎn)導(dǎo)中具有不同的功能從而形成一因多效。系統(tǒng)研究多效基因在各種逆境脅迫應(yīng)答中的具體功能及其對生長發(fā)育的影響對于未來鑒定作物廣譜抗逆調(diào)控因子具有重要作用。本研究通過生物信息學(xué)相關(guān)手段篩選出一批棉花響應(yīng)多逆境的重要候選基因,并對其中三個基因進(jìn)行了功能鑒定。取得的主要結(jié)果如下:1.利用STIFDB和GENEVESTIGATOR V3等數(shù)據(jù)庫,篩選出826個擬南芥廣譜響應(yīng)逆境的調(diào)節(jié)因子。通過同源序列法進(jìn)一步分離篩選出38個棉花廣譜響應(yīng)逆境的候選基因。對該38個棉花候選基因在鹽脅迫、冷脅迫以及ABA處理下的表達(dá)模式進(jìn)行了分析,有37個基因至少對一種處理有響應(yīng)。為了進(jìn)一步驗(yàn)證該策略的可靠性,我們選擇其中三個基因(GhATAF1、GhJAZ2、GhHB12)進(jìn)行了系統(tǒng)的功能研究。2.研究發(fā)現(xiàn)GhATAF1編碼一個有自激活活性的NAC轉(zhuǎn)錄因子,不僅受到ABA、鹽脅迫和冷脅迫等非生物脅迫的誘導(dǎo)表達(dá),還強(qiáng)烈受到MeJA、SA和棉花黃萎病菌V991的誘導(dǎo)表達(dá),正調(diào)控棉花的耐鹽性,同時(shí)負(fù)調(diào)控棉花對黃萎病菌的抗性。GhATAF1能通過激活棉花鉀離子/鈉離子共轉(zhuǎn)運(yùn)子GhHKT1的表達(dá),同時(shí)還能激活A(yù)BA信號關(guān)鍵應(yīng)答基因GhABI4,液泡氫離子-焦磷酸磷酸酶GhAVP1,晚期胚胎豐富蛋白GhLEA3,GhLEA6,干旱應(yīng)答基因GhRD22和GhDREB2A的表達(dá),減少棉花組織鈉離子的積累,減輕細(xì)胞的損傷,從而提高棉花的耐鹽性;此外我們發(fā)現(xiàn)GhATAF1能夠激活棉花水楊酸信號路徑,同時(shí)抑制茉莉酸信號路徑,最終削弱棉花對大麗輪枝菌和灰霉菌的抗性。3.研究發(fā)現(xiàn)GhJAZ2基因同時(shí)受到ABA、MeJA、SA、鹽脅迫、冷脅迫以及大麗輪枝菌V991的誘導(dǎo)上調(diào)表達(dá),是一個重要的調(diào)控棉花防御反應(yīng)和生長發(fā)育的節(jié)點(diǎn)基因。GhJAZ2作為一個棉花茉莉酸信號的抑制子,通過與GhJAZ12互作,形成異源二聚體,直接抑制GhMYC2-like和GhPR10,GhR1,GhD2等抗性相關(guān)蛋白,負(fù)調(diào)控棉花抗病性和抗蟲性;可能通過與DELLA蛋白的互作,間接激活赤霉素信號路徑,調(diào)控棉花花期、種子萌發(fā)和果枝夾角等生長發(fā)育過程。此外GhJAZ2還參與了高溫脅迫和低溫脅迫下棉花雄蕊的發(fā)育。4.研究發(fā)現(xiàn)GhHB12基因同時(shí)受到ABA、MeJA、SA、鹽脅迫、冷脅迫以及大麗輪枝菌V991的誘導(dǎo)上調(diào)表達(dá),是一個重要的調(diào)控棉花防御反應(yīng)和生長發(fā)育的多效基因。GhHB12負(fù)調(diào)控植物ABA信號路徑,負(fù)調(diào)控植物對非生物逆境的抗性;同時(shí)負(fù)調(diào)控棉花JA信號路徑,削弱棉花的抗病性和抗蟲性;此外GhHB12通過與GhTCP1互作,協(xié)同負(fù)調(diào)控生長素信號路徑,抑制棉花株高,促進(jìn)葉枝的發(fā)生;同時(shí)可以與GhSPLs互作,相互拮抗,直接調(diào)控GhFT、GhSOC1、GhFUL等開花基因的表達(dá),調(diào)控棉花花期、葉枝和果枝的發(fā)育。此外GhHB12還參與對棉花葉形的調(diào)控。我們還發(fā)現(xiàn)GhHB12受到光周期和節(jié)律的調(diào)控,GhHB12調(diào)控棉花株型的功能在短日照條件下會喪失。以上結(jié)果表明利用生物信息學(xué)分析能提高鑒定棉花響應(yīng)多重逆境關(guān)鍵調(diào)控因子的有效性,以上結(jié)果為棉花響應(yīng)多重逆境反應(yīng)的生理機(jī)制提供了參考。
[Abstract]:The plant can sense various environmental changes of the outside through the complex and fine control network, activate the relevant signal path, resist the stress of adversity, but at the same time, it is often accompanied by the change of the growth and development process and the yield. Due to the complexity of the gene regulation network, some genes are located at the intersection of multiple signal paths to form the hub gene and have different functions in multiple signal transduction to form a multi-effect. The specific function of the multi-effect gene in various stress-stress responses and its influence on the growth and development of the system have an important role in the future identification of crop broad-spectrum anti-reverse regulation factors. In this study, a number of important candidate genes in response to multiple adversity were selected by means of bioinformatics, and the three genes were identified. The main results are as follows: 1. By using the database of STIFDB and GENVESTIGATOR V3, the regulation factors of the broad-spectrum response of 826 arabidopsis thaliana were selected. and the candidate genes of the 38 cotton broad-spectrum response adversity are further separated and screened by the homologous sequence method. The expression patterns of 38 cotton candidate genes under salt stress, cold stress and ABA treatment were analyzed. To further verify the reliability of the strategy, we selected three of the genes (GhATAF1, GJAK2, GHB12) to perform the functional study of the system. It is found that GhATAF1 encodes a NAC transcription factor with self-activation activity, not only is induced by abiotic stress such as ABA, salt stress and cold stress, but also is strongly influenced by the induced expression of MeJA, SA and Verticillium dahliae V991, and the salt tolerance of cotton is being regulated. and meanwhile, the resistance of the cotton to the Verticillium dahliae is negatively regulated. The GhATAF1 can activate the expression of the cotton potassium ion/ sodium ion cotransporter GhHKT1, and can also activate the expression of the ABA signal key response gene GhABI4, the vacuole hydrogen ion-pyrophosphatase GhAVP1, the late embryo-rich protein GhLEA3, GhLEA6, the drought response gene GhR22 and the GhDREB2A, the accumulation of sodium ions in the cotton tissue is reduced, the damage of the cells is reduced, and the salt tolerance of the cotton is improved; in addition, the GhATAF1 is found to be capable of activating the path of the cotton salicylic acid signal, and meanwhile, the signal path of the jasmonic acid is inhibited, and the resistance of the cotton to the large-lily and the gray mold is finally weakened. It was found that the GhJAK2 gene was induced up-regulated by ABA, MeJA, SA, salt stress, cold stress and V991, which is an important node gene for regulating and controlling the defense response and growth of cotton. GhJAK2 is used as a suppressor of a cotton jasmonic acid signal, and through interaction with the GhJZ12, the heterogenous dimer is formed to directly inhibit the resistance-related proteins such as GhMYC2-like and GhPR10, GhR1, GGD2 and the like, and the resistance and the insect resistance of the cotton are negatively controlled; and the gibberellin signal path can be indirectly activated through the interaction with the DELLA protein, the growth and development process of controlling the flowering stage, the seed germination and the fruit branch included angle and the like of the cotton. In addition, GJAK2 is involved in the development of the stamen of the cotton under high-temperature stress and low-temperature stress. It was found that the GHB12 gene was induced up-regulated by ABA, MeJA, SA, salt stress, cold stress and V991, which is an important multi-effect gene for regulating and controlling the defense response and growth of cotton. the GhHB12 negative control plant ABA signal path and the negative control plant resistance to abiotic stress; and meanwhile, the cotton JA signal path is negatively regulated, the disease resistance and the insect resistance of the cotton are reduced, and the expression of the flowering genes such as the GhFT, the GhSOC1 and the GhFUL can be directly controlled, and the development of the flowering, branch and fruit branches of the cotton can be controlled. In addition, the GhHB12 is involved in the control of the leaf shape of the cotton. We also found that the GhHB12 was regulated by the photoperiod and the rhythm, and the function of the GhHB12 in the control of the cotton plant type was lost under short-day conditions. The above results show that bioinformatics analysis can improve the effectiveness of the key control factors for identifying the response of the cotton in response to the multiple adversity, and the above results provide a reference for the physiological mechanism of the response of the cotton to the multiple adversity.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S562
本文編號：2315565
[Abstract]:The plant can sense various environmental changes of the outside through the complex and fine control network, activate the relevant signal path, resist the stress of adversity, but at the same time, it is often accompanied by the change of the growth and development process and the yield. Due to the complexity of the gene regulation network, some genes are located at the intersection of multiple signal paths to form the hub gene and have different functions in multiple signal transduction to form a multi-effect. The specific function of the multi-effect gene in various stress-stress responses and its influence on the growth and development of the system have an important role in the future identification of crop broad-spectrum anti-reverse regulation factors. In this study, a number of important candidate genes in response to multiple adversity were selected by means of bioinformatics, and the three genes were identified. The main results are as follows: 1. By using the database of STIFDB and GENVESTIGATOR V3, the regulation factors of the broad-spectrum response of 826 arabidopsis thaliana were selected. and the candidate genes of the 38 cotton broad-spectrum response adversity are further separated and screened by the homologous sequence method. The expression patterns of 38 cotton candidate genes under salt stress, cold stress and ABA treatment were analyzed. To further verify the reliability of the strategy, we selected three of the genes (GhATAF1, GJAK2, GHB12) to perform the functional study of the system. It is found that GhATAF1 encodes a NAC transcription factor with self-activation activity, not only is induced by abiotic stress such as ABA, salt stress and cold stress, but also is strongly influenced by the induced expression of MeJA, SA and Verticillium dahliae V991, and the salt tolerance of cotton is being regulated. and meanwhile, the resistance of the cotton to the Verticillium dahliae is negatively regulated. The GhATAF1 can activate the expression of the cotton potassium ion/ sodium ion cotransporter GhHKT1, and can also activate the expression of the ABA signal key response gene GhABI4, the vacuole hydrogen ion-pyrophosphatase GhAVP1, the late embryo-rich protein GhLEA3, GhLEA6, the drought response gene GhR22 and the GhDREB2A, the accumulation of sodium ions in the cotton tissue is reduced, the damage of the cells is reduced, and the salt tolerance of the cotton is improved; in addition, the GhATAF1 is found to be capable of activating the path of the cotton salicylic acid signal, and meanwhile, the signal path of the jasmonic acid is inhibited, and the resistance of the cotton to the large-lily and the gray mold is finally weakened. It was found that the GhJAK2 gene was induced up-regulated by ABA, MeJA, SA, salt stress, cold stress and V991, which is an important node gene for regulating and controlling the defense response and growth of cotton. GhJAK2 is used as a suppressor of a cotton jasmonic acid signal, and through interaction with the GhJZ12, the heterogenous dimer is formed to directly inhibit the resistance-related proteins such as GhMYC2-like and GhPR10, GhR1, GGD2 and the like, and the resistance and the insect resistance of the cotton are negatively controlled; and the gibberellin signal path can be indirectly activated through the interaction with the DELLA protein, the growth and development process of controlling the flowering stage, the seed germination and the fruit branch included angle and the like of the cotton. In addition, GJAK2 is involved in the development of the stamen of the cotton under high-temperature stress and low-temperature stress. It was found that the GHB12 gene was induced up-regulated by ABA, MeJA, SA, salt stress, cold stress and V991, which is an important multi-effect gene for regulating and controlling the defense response and growth of cotton. the GhHB12 negative control plant ABA signal path and the negative control plant resistance to abiotic stress; and meanwhile, the cotton JA signal path is negatively regulated, the disease resistance and the insect resistance of the cotton are reduced, and the expression of the flowering genes such as the GhFT, the GhSOC1 and the GhFUL can be directly controlled, and the development of the flowering, branch and fruit branches of the cotton can be controlled. In addition, the GhHB12 is involved in the control of the leaf shape of the cotton. We also found that the GhHB12 was regulated by the photoperiod and the rhythm, and the function of the GhHB12 in the control of the cotton plant type was lost under short-day conditions. The above results show that bioinformatics analysis can improve the effectiveness of the key control factors for identifying the response of the cotton in response to the multiple adversity, and the above results provide a reference for the physiological mechanism of the response of the cotton to the multiple adversity.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S562
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 徐理;朱龍付;張獻(xiàn)龍;;棉花抗黃萎病機(jī)制研究進(jìn)展[J];作物學(xué)報(bào);2012年09期
2 朱龍付;涂禮莉;曾范昌;劉迪秋;張獻(xiàn)龍;;一種適合于cDNA文庫構(gòu)建的高質(zhì)量棉花RNA的簡單抽提法(英文)[J];作物學(xué)報(bào);2005年12期
相關(guān)博士學(xué)位論文 前5條
1 李超;棉花GbWRKY1在植物發(fā)育及防御反應(yīng)中的功能分析[D];華中農(nóng)業(yè)大學(xué);2014年
2 徐飛;落葉型棉花黃萎病菌在華中棉區(qū)廣泛分布及其原因分析[D];華中農(nóng)業(yè)大學(xué);2012年
3 徐理;棉花與黃萎病菌的分子互作機(jī)制研究及GbWRKY1基因的功能鑒定[D];華中農(nóng)業(yè)大學(xué);2011年
4 陳捷胤;海島棉LRR-TM類抗病基因GbaVd1和GbaVd2的克隆與功能研究[D];中國農(nóng)業(yè)科學(xué)院;2010年
5 金雙俠;棉花遺傳轉(zhuǎn)化體系的優(yōu)化及突變體的創(chuàng)制[D];華中農(nóng)業(yè)大學(xué);2006年
相關(guān)碩士學(xué)位論文 前1條
1 劉琳琳;番茄Vel基因在棉花中的功能驗(yàn)證[D];華中農(nóng)業(yè)大學(xué);2013年
，本文編號：2315565
本文鏈接：http://www.sikaile.net/shoufeilunwen/nykjbs/2315565.html
最近更新
教材專著
