對比根系轉(zhuǎn)錄組研究棉花鹽脅迫應(yīng)答及耐受機(jī)理
發(fā)布時(shí)間：2018-10-21 07:41
【摘要】：土壤鹽漬化是限制農(nóng)作物生產(chǎn)的全球性非生物脅迫因素之一,土壤飽和浸提液電導(dǎo)率等于4.0dS m-1的土壤即為鹽漬化土壤(相當(dāng)于40mM NaC1)。提高農(nóng)作物的耐鹽抗旱能力已成為現(xiàn)代農(nóng)業(yè)生產(chǎn)中一個(gè)亟待解決的問題。陸地棉(Gossypium hirsutum L.)作為全球種植面積最大的纖維作物,在飽和浸提液電導(dǎo)率大于7.7dS m-1的鹽漬化土壤中才開始表現(xiàn)出減產(chǎn),具有較高的耐鹽能力,被視為開發(fā)利用鹽堿地的“先鋒作物”以及研究植物耐鹽機(jī)制的模式植物。因此,棉花耐鹽分子機(jī)理的研究受到廣泛的重視。 本研究選擇一個(gè)相對敏鹽陸地棉材料“中G5”,構(gòu)建了其三葉一心期幼苗根系的7個(gè)全長cDNA文庫,包括150mM NaCl鹽脅迫后3h、12h和48h三個(gè)時(shí)間點(diǎn)的處理文庫,以及3個(gè)對應(yīng)時(shí)間點(diǎn)的對照文庫,另加一個(gè)0h時(shí)間點(diǎn)對照文庫。 從7個(gè)文庫中隨機(jī)挑選約3,300個(gè)克隆進(jìn)行5’端焦磷酸測序,去掉載體接頭及低質(zhì)量序列后共獲得20,358條高質(zhì)量ESTs；利用CAP3拼接后得到8,516條niESTs,包括2,914個(gè)contigs和5,602個(gè)singletons。比對最新拼接棉花uniESTs結(jié)果發(fā)現(xiàn)1,795條uniESTs為本研究首次報(bào)道,另外6,721個(gè)τmiESTs (78.9%)有同源結(jié)果。 利用BLAST2GO對8,516條uniESTs進(jìn)行了GO注釋后,對比分析了三個(gè)時(shí)間點(diǎn)對照和脅迫文庫中uniESTs的GO變化,發(fā)現(xiàn)鹽脅迫后棉花根系激活了一系列應(yīng)答調(diào)控進(jìn)程,如生長調(diào)控、刺激應(yīng)答、脅迫信號傳導(dǎo)以及轉(zhuǎn)錄調(diào)控等以應(yīng)答鹽脅迫�；趯Ρ葘φ蘸兔{迫文庫間轉(zhuǎn)錄因子ESTs發(fā)現(xiàn),MYB、MYB-related、WRKY、bHLH、GRAS和ERF家族轉(zhuǎn)錄因子在鹽脅迫后3h顯著富集,而NAC家族轉(zhuǎn)錄因子則在鹽脅迫后12和48h顯著富集。 Fish er統(tǒng)計(jì)檢驗(yàn)發(fā)現(xiàn)65個(gè)鹽脅迫應(yīng)答顯著調(diào)控基因,主要功能涉及活性氧清除,赤霉素代謝,脅迫應(yīng)答,水分及其他物質(zhì)的跨膜轉(zhuǎn)運(yùn),信號傳導(dǎo)以及轉(zhuǎn)錄調(diào)控等。熒光定量PCR檢測支持了Fis1ler檢驗(yàn)的結(jié)果,并且在酵母中超表達(dá)部分差異應(yīng)答基因顯著提高了酵母的耐鹽能力。GO和DEGs的分析結(jié)果均表明,ROS以及GAs信號通路積極參與了棉花的鹽脅迫應(yīng)答調(diào)控,結(jié)合模式植物中ROS與GAs間互作代謝的分析結(jié)果,我們提出一個(gè)ROS-GAs信號互作參與棉花鹽脅迫應(yīng)答的調(diào)控模型。 通過研究鹽脅迫后多個(gè)時(shí)間點(diǎn)棉花根系的轉(zhuǎn)錄組應(yīng)答調(diào)控,首次揭示了棉花鹽脅迫動態(tài)的應(yīng)答過程及多信號互作調(diào)控的耐鹽應(yīng)答機(jī)理,鑒定了一批具有耐鹽功能的棉花基因。本實(shí)驗(yàn)構(gòu)建的全長cDNA文庫及隨機(jī)測序獲得的ESTs序列信息為克隆棉花基因提供了便利,并為進(jìn)一步研究棉花耐鹽機(jī)制、遺傳改良棉花耐鹽性提供了基因資源。
[Abstract]:Soil salinization is one of the global abiotic stress factors that limit crop production. Soil whose conductivity of saturated extract is equal to 4.0dS m-1 is salinized soil (equivalent to 40mM NaC1). Improving the salt tolerance and drought resistance of crops has become an urgent problem in modern agricultural production. Upland cotton (Gossypium hirsutum L.) As the largest fiber crop planted in the world, it is only in salinized soil where the conductivity of saturated extract is greater than 7.7dS m-1 that it begins to show reduced production and has higher salt tolerance. It is regarded as a pioneer crop and a model plant to study the mechanism of salt tolerance in saline-alkali land. Therefore, the molecular mechanism of salt tolerance in cotton has received extensive attention. In this study, 7 full-length cDNA libraries were constructed from the root system of three leaf seedlings of a relatively salt-sensitive upland cotton cultivar "ZhongG5", including three treatment libraries at 3 h, 12 h and 48 h after 150mM NaCl salt stress. And 3 reference libraries corresponding to time points, plus a 0 h time point library. About 3300 clones were randomly selected from 7 libraries for 5 'terminal pyrosequencing. 20358 high quality ESTs; were obtained after removing the vector connector and low quality sequence. 8516 niESTs, including 2914 contigs and 5602 singletons. were obtained by using CAP3 splicing. Compared with the latest cotton uniESTs results, 1795 uniESTs were reported for the first time, and 6721 蟿 miESTs (78.9%) had homologous results. After 8516 uniESTs were annotated with GO by BLAST2GO, the changes of uniESTs GO in three time points control and stress library were compared and analyzed. It was found that cotton root system activated a series of response regulation processes, such as growth regulation and stimulative response, after salt stress. Stress signal transduction and transcriptional regulation in response to salt stress. MYB,MYB-related,WRKY,bHLH,GRAS and ERF family transcription factors were significantly enriched at 3 h after salt stress based on contrastive control and stress interlibrary transcription factor (ESTs). On the other hand, NAC family transcription factors were significantly enriched in. Fish er at 12 and 48 hours after salt stress. 65 genes were found to regulate salt stress response, and their main functions were active oxygen scavenging, gibberellin metabolism and stress response. Transmembrane transport, signal transduction and transcriptional regulation of water and other substances. Fluorescence quantitative PCR test supports the results of Fis1ler test. The results of GO and DEGs analysis showed that ROS and GAs signaling pathway were actively involved in the regulation of salt stress response in cotton. Based on the analysis of the interaction metabolism between ROS and GAs in model plants, we proposed a model for regulating the interaction of ROS-GAs signals in cotton salt stress response. By studying the transcriptional response regulation of cotton roots at different time points after salt stress, the dynamic response process of cotton salt stress and the mechanism of salt tolerance under the regulation of multi-signal interaction were revealed for the first time, and a number of salt tolerant genes were identified. The full-length cDNA library constructed in this study and the ESTs sequence information obtained by random sequencing provided genetic resources for cloning cotton gene and further studying the mechanism of cotton salt tolerance and genetic improvement of cotton salt tolerance.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S562
本文編號：2284415
[Abstract]:Soil salinization is one of the global abiotic stress factors that limit crop production. Soil whose conductivity of saturated extract is equal to 4.0dS m-1 is salinized soil (equivalent to 40mM NaC1). Improving the salt tolerance and drought resistance of crops has become an urgent problem in modern agricultural production. Upland cotton (Gossypium hirsutum L.) As the largest fiber crop planted in the world, it is only in salinized soil where the conductivity of saturated extract is greater than 7.7dS m-1 that it begins to show reduced production and has higher salt tolerance. It is regarded as a pioneer crop and a model plant to study the mechanism of salt tolerance in saline-alkali land. Therefore, the molecular mechanism of salt tolerance in cotton has received extensive attention. In this study, 7 full-length cDNA libraries were constructed from the root system of three leaf seedlings of a relatively salt-sensitive upland cotton cultivar "ZhongG5", including three treatment libraries at 3 h, 12 h and 48 h after 150mM NaCl salt stress. And 3 reference libraries corresponding to time points, plus a 0 h time point library. About 3300 clones were randomly selected from 7 libraries for 5 'terminal pyrosequencing. 20358 high quality ESTs; were obtained after removing the vector connector and low quality sequence. 8516 niESTs, including 2914 contigs and 5602 singletons. were obtained by using CAP3 splicing. Compared with the latest cotton uniESTs results, 1795 uniESTs were reported for the first time, and 6721 蟿 miESTs (78.9%) had homologous results. After 8516 uniESTs were annotated with GO by BLAST2GO, the changes of uniESTs GO in three time points control and stress library were compared and analyzed. It was found that cotton root system activated a series of response regulation processes, such as growth regulation and stimulative response, after salt stress. Stress signal transduction and transcriptional regulation in response to salt stress. MYB,MYB-related,WRKY,bHLH,GRAS and ERF family transcription factors were significantly enriched at 3 h after salt stress based on contrastive control and stress interlibrary transcription factor (ESTs). On the other hand, NAC family transcription factors were significantly enriched in. Fish er at 12 and 48 hours after salt stress. 65 genes were found to regulate salt stress response, and their main functions were active oxygen scavenging, gibberellin metabolism and stress response. Transmembrane transport, signal transduction and transcriptional regulation of water and other substances. Fluorescence quantitative PCR test supports the results of Fis1ler test. The results of GO and DEGs analysis showed that ROS and GAs signaling pathway were actively involved in the regulation of salt stress response in cotton. Based on the analysis of the interaction metabolism between ROS and GAs in model plants, we proposed a model for regulating the interaction of ROS-GAs signals in cotton salt stress response. By studying the transcriptional response regulation of cotton roots at different time points after salt stress, the dynamic response process of cotton salt stress and the mechanism of salt tolerance under the regulation of multi-signal interaction were revealed for the first time, and a number of salt tolerant genes were identified. The full-length cDNA library constructed in this study and the ESTs sequence information obtained by random sequencing provided genetic resources for cloning cotton gene and further studying the mechanism of cotton salt tolerance and genetic improvement of cotton salt tolerance.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S562
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 師恭曜;王玉美;華金平;;水通道蛋白與高等植物的耐鹽性[J];中國農(nóng)業(yè)科技導(dǎo)報(bào);2012年04期
2 張國偉;路海玲;張雷;陳兵林;周治國;;棉花萌發(fā)期和苗期耐鹽性評價(jià)及耐鹽指標(biāo)篩選[J];應(yīng)用生態(tài)學(xué)報(bào);2011年08期
，本文編號：2284415
本文鏈接：http://sikaile.net/shoufeilunwen/nykjbs/2284415.html
最近更新
教材專著
