本文選題：茶樹(shù)　切入點(diǎn)：遺傳變異　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：鋁毒是酸性土壤中植物生長(zhǎng)和作物生產(chǎn)的主要限制因素。茶樹(shù)起源于我國(guó)西南地區(qū),在亞熱帶、溫帶濕潤(rùn)的氣候條件下,形成了“喜酸喜鋁怕堿”的特性,其生長(zhǎng)最適宜土壤pH為4.5-5.5,其鋁含量是其它植物物種的幾十倍,且沒(méi)有任何鋁毒害癥狀,表現(xiàn)出鋁超富集的特性,表明茶樹(shù)具有耐鋁和鋁富集機(jī)制,因此,明確茶樹(shù)的耐鋁和鋁富集的機(jī)制,對(duì)于培育具有耐酸耐鋁性的農(nóng)作物、提高農(nóng)作物產(chǎn)量及促進(jìn)農(nóng)業(yè)可持續(xù)發(fā)展具有非常重要意義。本試驗(yàn)明確了37份不同茶樹(shù)基因型嫩梢的鋁富集及根系生長(zhǎng)響應(yīng)不同鋁濃度的遺傳變異,挖掘了嘉茗1號(hào)茶樹(shù)幼苗根系響應(yīng)不同鋁濃度(0 mM、0.2 mM、1.0 mM)的鋁富集候選基因,分析了C2H2型鋅指結(jié)構(gòu)轉(zhuǎn)錄因子CsSTOP1在49份茶樹(shù)基因型中的序列變異,結(jié)果如下:1.本試驗(yàn)采用營(yíng)養(yǎng)液培養(yǎng)系統(tǒng),設(shè)置0、0.2、0.4 mM Al濃度梯度,培養(yǎng)4個(gè)月后,測(cè)定了37份茶樹(shù)基因型的鋁濃度及根系生長(zhǎng)的遺傳變異。在0 m M Al濃度條件下,僅有福云6號(hào)、龍井43號(hào)、黔湄701、金牡丹4個(gè)基因型長(zhǎng)出白色吸收根,占10.81%;在0.2 mM Al濃度條件下,24個(gè)茶樹(shù)基因型長(zhǎng)出白色新吸收根,占64.86%;在0.4 m M Al條件下,28個(gè)基因型長(zhǎng)出白色新吸收根,占75.68%。結(jié)果表明,在適宜鋁濃度范圍內(nèi),鋁促進(jìn)茶樹(shù)根系生長(zhǎng),0.4 mM Al促進(jìn)作用大于0.2 mM Al。與不加鋁相比,0.4 mM Al濃度處理后,25個(gè)茶樹(shù)基因型的嫩梢鋁濃度均有不同程度的增加,表明不同茶樹(shù)基因型具有不同的鋁吸收或分配存在廣泛的遺傳變異。2.為了挖掘與茶樹(shù)耐鋁或聚鋁相關(guān)的候選基因,采用轉(zhuǎn)錄組對(duì)在0 mM、0.2mM、1.0 m M Al濃度處理?xiàng)l件下的嘉茗1號(hào)茶樹(shù)幼苗根系進(jìn)行轉(zhuǎn)錄組測(cè)序分析,共獲得70.18 GB原始?jí)A基序列數(shù)據(jù),共組裝了213699個(gè)高質(zhì)量單基因簇(unigenes)。篩選得到不同類(lèi)型的差異表達(dá)基因,主要包括轉(zhuǎn)錄因子、轉(zhuǎn)運(yùn)蛋白、氧化應(yīng)激通路蛋白,以及涉及多糖和細(xì)胞壁代謝、能量和次級(jí)代謝、有機(jī)酸陰離子和酶分泌、三羧酸循環(huán)(TCA)調(diào)控、信號(hào)傳導(dǎo)和生長(zhǎng)激素生物合成代謝途徑等基因,以上基因可能是茶樹(shù)鋁富集潛在候選基因。3.比對(duì)分析了49個(gè)茶樹(shù)基因型的CsSTOP1基因序列,發(fā)現(xiàn)了63個(gè)單核苷酸(SNP)變異位點(diǎn),其中33個(gè)位點(diǎn)的變異導(dǎo)致氨基酸發(fā)生改變,30個(gè)為同義變異,表明不同茶樹(shù)基因型間CsSTOP1基因遺傳變異豐富,還發(fā)現(xiàn)在5個(gè)茶樹(shù)基因型(云南地方群體良種、云南大葉種、云抗27號(hào)、云茶普蕊、勐庫(kù)大葉種)中CsSTOP1出現(xiàn)3個(gè)氨基酸序列的缺失,屬于阿薩姆茶[C.sinensis var.Assamica(Masters)Kitamura]變種,該基因可能參與茶樹(shù)耐鋁聚鋁的調(diào)控,這3個(gè)氨基酸的變異可能與茶樹(shù)鋁富集存在重要聯(lián)系。
[Abstract]:Aluminum toxicity is the main limiting factor for plant growth and crop production in acidic soils. Tea trees originated in southwest China and formed "acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-acid-@@. The optimum soil pH was 4.5-5.5, and its aluminum content was several times higher than that of other plant species. Moreover, it had no symptoms of aluminum toxicity and showed the characteristics of aluminum superenrichment, which indicated that tea plants had the mechanism of aluminum tolerance and aluminum enrichment. To clarify the mechanism of aluminum tolerance and aluminum enrichment of tea plants, for the cultivation of acid- and aluminum-tolerant crops, It is very important to increase crop yield and promote the sustainable development of agriculture. In this experiment, the accumulation of aluminum in 37 tea genotypes and the genetic variation of different Al concentrations in response to root growth were determined. The candidate genes for aluminum enrichment in roots of Jiaming No. 1 tea seedlings responding to different aluminum concentrations (0 mm ~ 0 mm ~ (2) m ~ (2)) and 1.0 mm / m ~ (-1) were excavated. The sequence variation of zinc finger structure transcription factor (CsSTOP1) of C _ 2H _ 2 type in 49 tea genotypes was analyzed. The results were as follows: 1.The concentration gradient of 0.4 mm Al was set up in a nutrient solution culture system. After 4 months of culture, the Al concentration of 37 tea genotypes and the genetic variation of root growth were determined. Under the condition of 0 mm Al concentration, Only 4 genotypes of Fuyun 6, Longjing 43, Qianmai 701, Golden Peony grew white absorbing roots, accounting for 10.81%, and 24 tea genotypes grew white new absorbing roots under 0.2 mm Al concentration. Under the condition of 0.4 mm Al, 28 genotypes grew white new absorbing roots, accounting for 75.68%. The results showed that, in the suitable concentration range of aluminum, The effects of Al on root growth of tea plants were more than 0.2 mm Al. The concentrations of Al in the young shoots of 25 tea genotypes increased in varying degrees compared with those without Al addition. The results indicate that different tea genotypes have different aluminum uptake or distribution. 2. In order to mine candidate genes related to aluminum tolerance or polyaluminum tolerance in tea plants, The root system of tea seedling of Jiaming No. 1 was analyzed by transcriptome sequencing under the concentration of 0 mMM 0.2mMM + 1.0 mm Al, and 70.18GB primordial sequence data were obtained. A total of 213699 high quality single gene clusters were assembled. Different types of differentially expressed genes were screened, including transcription factors, transporters, oxidative stress pathway proteins, polysaccharide and cell wall metabolism, energy and secondary metabolism. Organic acid anion and enzyme secretion, tricarboxylic acid cycle TCA regulation, signal transduction and growth hormone biosynthesis pathway, The above genes may be potential candidate genes for aluminum enrichment in tea plants. The CsSTOP1 gene sequences of 49 tea genotypes were compared and 63 SNPs mutation sites were found. Among them, 33 loci caused amino acid changes and 30 were synonymous, indicating that the genetic variation of CsSTOP1 gene was abundant among different tea genotypes, and 5 tea genotypes (Yunnan local population, Yunnan large leaf species) were also found. There were three amino acid sequence deletions of CsSTOP1 in Yunkang 27, Yuncha Rui, Mengku leaf species), which belong to C. sinensis var.Assamica(Masters)Kitamura variety, and this gene may be involved in the regulation of aluminum polyaluminum tolerance in tea plants. The variation of these three amino acids may be related to aluminum enrichment in tea plants.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S571.1

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