本文選題：亞精胺合成酶(SPDS) + 馬鈴薯　； 參考：《湖南農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：馬鈴薯品種逆境抗性的改善,是馬鈴薯育種的主要目標(biāo)之一,多胺類(lèi)物質(zhì)與植物逆境抗性關(guān)系密切,植物亞精胺合成酶是多胺生物合成代謝中的關(guān)鍵酶之一。本研究以轉(zhuǎn)植物亞精胺合成酶基因SPDS的馬鈴薯植株為材料,對(duì)轉(zhuǎn)SPDS馬鈴薯植株導(dǎo)入基因穩(wěn)定性和表達(dá)情況,低溫脅迫期間轉(zhuǎn)SPDS馬鈴薯植株的低溫響應(yīng)表現(xiàn)、冷凍處理葉片相對(duì)電導(dǎo)率、多胺含量、生理抗性指標(biāo),低溫脅迫遭遇前后的轉(zhuǎn)錄組測(cè)序的差異表達(dá)基因進(jìn)行了檢測(cè)、觀察和初步的生物信息學(xué)分析。主要研究結(jié)果如下：1、對(duì)14個(gè)轉(zhuǎn)SPDS馬鈴薯株系的PCR、RT-PCR檢測(cè)和多胺HPLC定量分析,擇優(yōu)篩選、初步獲得6個(gè)導(dǎo)入基因遺傳及表達(dá)穩(wěn)定、多胺代謝強(qiáng)化顯著的轉(zhuǎn)SPDS馬鈴薯株系。2、初步篩選的6個(gè)轉(zhuǎn)SPDS馬鈴薯株系的低溫脅迫應(yīng)答響應(yīng)、低溫脅迫處理后回復(fù)常溫生育狀況、人工冷凍低溫處理葉片相對(duì)電導(dǎo)率三方面的變化均優(yōu)于對(duì)照植株,說(shuō)明SPDS的導(dǎo)入提高了馬鈴薯植株的低溫抗性。3、遭遇低溫脅迫轉(zhuǎn)SPDS馬鈴薯植株的多胺含量、光合作用和生理抗性指標(biāo)的變化均優(yōu)于對(duì)照種,從生理水平對(duì)轉(zhuǎn)SPDS馬鈴薯植株低溫抗性改善的機(jī)理機(jī)制進(jìn)行了初步探討。4、以轉(zhuǎn)SPDS馬鈴薯株系02-42及其對(duì)照種東農(nóng)303為材料,運(yùn)用DGE技術(shù)對(duì)低溫脅迫遭遇前后馬鈴薯植株的表達(dá)差異基因進(jìn)行了初步的研究�；趯�(duì)照和轉(zhuǎn)SPDS植株所特有的差異表達(dá)基因的比較分析,初步發(fā)現(xiàn)有14個(gè)表達(dá)差異達(dá)顯著水平的基因,其中明顯上調(diào)的7個(gè)基因有利于轉(zhuǎn)SPDS馬鈴薯植株低溫抗性改善。
[Abstract]:The improvement of stress resistance of potato varieties is one of the main objectives of potato breeding. Polyamines are closely related to plant stress resistance. Spermidine synthase is one of the key enzymes in polyamine biosynthesis.In this study, the transgenic potato plants with spermatine synthase gene SPDS were used as materials to study the gene stability and expression of SPDS transgenic potato plants, and the low temperature response of the transgenic potato plants to SPDS during low temperature stress.The relative electrical conductivity, polyamine content, physiological resistance index and differentially expressed genes of transcriptome sequencing before and after cold stress were detected, observed and preliminarily analyzed by bioinformatics.The main results were as follows: 1. PCR and polyamine HPLC quantitative analysis were performed on 14 potato lines transferred to SPDS, and 6 introduced genes were selected for genetic analysis and stable expression.The polyamine metabolism was enhanced significantly in SPDS potato strain. 2. The response to low temperature stress of 6 SPDS potato lines was preliminarily screened, and the growth status at room temperature was restored after low temperature stress treatment.The changes of relative electrical conductivity in leaves of artificial cryopreservation treatment were better than that of control plants, indicating that the introduction of SPDS increased the low temperature resistance of potato plants, and the polyamines content of potato plants transferred to SPDS under low temperature stress.The changes of photosynthesis and physiological resistance indexes were better than that of the control species. The mechanism of improving the low temperature resistance of transgenic SPDS potato plants was preliminarily discussed from physiological level. The transgenic potato strain 02-42 and its control Dongnong 303 were used as materials.DGE technique was used to study the differentially expressed genes in potato plants before and after exposure to low temperature stress.Based on the comparative analysis of the specific differentially expressed genes between the control and transgenic SPDS plants, 14 genes with significant difference were found, among which 7 genes upregulated were beneficial to the improvement of low temperature resistance of transgenic potato plants with SPDS.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S532

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