發(fā)布時(shí)間：2018-03-31 04:35

  本文選題：‘云香’水仙　切入點(diǎn)：基因克隆　出處：《西北植物學(xué)報(bào)》2017年02期

【摘要】：為探究多花水仙ACS基因的序列特征及功能,以‘云香’水仙盛花期花瓣為試驗(yàn)材料,根據(jù)‘云香’水仙花朵轉(zhuǎn)錄組數(shù)據(jù)信息,通過RT-PCR方法克隆出1個(gè)ACS基因,命名為NtACS1(GenBank KX082936);NtACS1開放閱讀框(ORF)長(zhǎng)度為552bp,編碼183個(gè)氨基酸。編碼蛋白質(zhì)分子量約為20.6KDa,理論等電點(diǎn)為6.30,不穩(wěn)定系數(shù)為65.49,屬于不穩(wěn)定的疏水性蛋白。通過qRT-PCR對(duì)‘云香’水仙不同時(shí)期花瓣和副冠中的NtACS1基因進(jìn)行了表達(dá)分析,得到與‘云香’水仙花朵轉(zhuǎn)錄組數(shù)據(jù)中相同的結(jié)果:NtACS1基因在‘云香’水仙花瓣和副冠中的表達(dá)都是隨著花衰老過程呈現(xiàn)逐漸下降的趨勢(shì),且NtACS1基因在花瓣和副冠中的表達(dá)峰值都在花苞期,表明NtACS1基因編碼的蛋白是在乙烯生物合成途徑的系統(tǒng)1發(fā)揮催化作用的ACC合成酶。成功構(gòu)建了NtACS1基因的正義植物表達(dá)載體,并通過農(nóng)桿菌介導(dǎo)法獲得8株轉(zhuǎn)基因煙草,PCR和RT-PCR檢測(cè)顯示其中有6株為陽性植株,初步證實(shí)NtACS1基因已導(dǎo)入煙草基因組中且在煙草中已表達(dá)。該研究結(jié)果為進(jìn)一步分析NtACS1基因的功能和后續(xù)轉(zhuǎn)化水仙延長(zhǎng)其花期研究奠定了基礎(chǔ)。
[Abstract]:In order to study the sequence and function of ACS gene of Narcissus multiflorum, a ACS gene was cloned by RT-PCR from petals of Narcissus in flowering period. Named NtACS1(GenBank KX082936NtACS1, the length of ORF is 552 BP, encoding 183 amino acids. The molecular weight of encoded protein is about 20.6KDa. the theoretical isoelectric point is 6.30, and the instability coefficient is 65.49. it is an unstable hydrophobic protein. The expression of NtACS1 gene in petal and accessory at different stages was analyzed. The same results were obtained from the transcriptional data of 'Yunxiang' Narcissus flowers. The expression of the gene in the petals and accessory crowns of 'Yunxiang' narcissus showed a decreasing trend with the flower senescence. The peak expression of NtACS1 gene in petal and accessory was in flower bud, indicating that the protein encoded by NtACS1 gene was a ACC synthase that played a catalytic role in ethylene biosynthesis pathway system 1. The sense plant expression vector of NtACS1 gene was successfully constructed. By Agrobacterium tumefaciens mediated PCR and RT-PCR analysis, 6 of the 8 transgenic tobacco plants were found to be positive. It was confirmed that the NtACS1 gene had been introduced into the tobacco genome and expressed in tobacco. The results laid a foundation for the further analysis of the function of NtACS1 gene and the subsequent transformation of narcissus to prolong its flowering period.
【作者單位】： 福建農(nóng)林大學(xué)園藝學(xué)院;福建農(nóng)林大學(xué)園藝植物遺傳育種研究所;廈門大學(xué)環(huán)境與生態(tài)學(xué)院;
【基金】：福建省種業(yè)創(chuàng)新與產(chǎn)業(yè)化工程項(xiàng)目(K8114001B)
【分類號(hào)】：Q943.2;S682.21
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本文編號(hào)：1689201