本文選題：大葉落地生根 + S-腺苷-L-高半胱氨酸水解酶基因(SAHH)　； 參考：《農(nóng)業(yè)生物技術(shù)學報》2016年04期

【摘要】：S-腺苷-L-高半胱氨酸水解酶(S-adenosyl-L-homocysteine hydrolase,SAHH)是已知的唯一能夠水解S-腺苷高半胱氨酸(S-adenosylhomocysteine,SAH)成半胱氨酸和腺苷的酶。SAH作為轉(zhuǎn)甲基反應的抑制劑,與S-腺苷甲硫氨酸(S-adenosyl-L-methionine,SAM,體內(nèi)甲基供體)的比值常用來作為衡量植物整體甲基化水平的重要指標。為鑒定大葉落地生根(Kalanchoe daigremontiana)SAHH的功能,本研究根據(jù)已知的大葉落地生根SAHH基因(Kd SAHH)序列(Gen Bank登錄號:KF953475)設計引物,對其進行干旱脅迫下的表達分析、亞細胞定位分析以及煙草(Nicotiana tabacum)轉(zhuǎn)化,并對T1代苗用20%PEG6000模擬干旱處理,測定0、12和24 h的相對含水量(relative water content,RWC)、葉綠素(chlorophyll,Chl)含量、丙二醛(malondialdehyde,MDA)含量、過氧化物酶(peroxidase,POD)含量、超氧化物歧化酶(superoxide dismutase,SOD)含量等與抗旱相關(guān)的生理指標。結(jié)果表明,Kd SAHH基因在輕度(20 d)、中度(35 d)、重度(50 d)干旱脅迫下表達量依次升高,表明該基因與抗旱性相關(guān);亞細胞定位結(jié)果顯示,該蛋白定位于細胞質(zhì)與細胞核;對轉(zhuǎn)基因煙草進行PCR檢測,獲得9株陽性植株;測定干旱脅迫下的相關(guān)生理指標,結(jié)果顯示,與野生型煙草相比,轉(zhuǎn)Kd SAHH煙草在干旱脅迫下能夠保持相對穩(wěn)定的RWC、Chl含量以及相對較低的MDA、POD含量。與野生型相比,轉(zhuǎn)Kd SAHH煙草受傷害輕,抗旱性增強,表明Kd SAHH基因能夠提高煙草植株的抗旱性。研究結(jié)果豐富了Kd SAHH基因的功能,為后續(xù)研究該基因提供了借鑒和參考。
[Abstract]:S-adenosyl-L-homocysteine hydrolase (S-adenosyl-L-homocysteine hydrolase SAHH) is the only known enzyme that can hydrolyze S-adenosylhomocysteine to cysteine and adenosylhomocysteine SAH as an inhibitor of transmethylation. The ratio of S-adenosyl-L-methionine (SAM) to S-adenosyl-L-methionine (S-adenosyl-L-methionine) is often used as an important index to evaluate the overall methylation level of plants. In order to identify the function of Kalanchoe daigremontiana)SAHH, primer was designed according to the known sequence of SAHH gene (Kd SAHH), Genin Bank accession number: KF953475), and the expression of KF953475 was analyzed under drought stress. Subcellular localization analysis and tobacco Nicotiana tabacum) transformation were performed. The relative water content, chlorophyll chlorophyllChl, malondialdehyde (malondialdehyde) and peroxidase (POD) contents in T1 generation seedlings were determined by 20%PEG6000 simulated drought treatment at 12 and 24 h, respectively. Superoxide dismutase (SOD) content and other physiological indexes related to drought resistance. The results showed that the expression of Kd SAHH gene increased under drought stress at 20 d, 35 d, 50 d, respectively, indicating that the gene was related to drought resistance, and the subcellular localization showed that the protein was located in cytoplasm and nucleus. Nine positive plants were obtained by PCR detection of transgenic tobacco, and the relative physiological indexes of transgenic tobacco under drought stress were determined. The results showed that compared with wild type tobacco, Under drought stress, the transgenic Kd SAHH tobacco could maintain a relatively stable content of RWCN Chl and a relatively low content of MDA-POD. Compared with wild type, transgenic Kd SAHH tobacco was less damaged and more drought-resistant, indicating that KD SAHH gene could improve the drought resistance of tobacco plants. The results enrich the function of KD SAHH gene and provide reference for further study of KD SAHH gene.
【作者單位】： 北京林業(yè)大學;
【基金】：國家高技術(shù)研究發(fā)展計劃(863計劃)項目(No.2013AA102607)
【分類號】：S682.33

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