本文選題：半夏 + 根癌農(nóng)桿菌��； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：半夏(Pinellia ternata(Thunb.)Breit)為天南星科半夏屬植物,是傳統(tǒng)名貴中藥材。半夏塊莖是中醫(yī)化痰燥濕、止痛消腫、止吐降逆的良藥。半夏在全國(guó)各地均有分布,雖分布廣泛,但由于過(guò)度開采和缺乏科學(xué)規(guī)范的種植技術(shù),半夏優(yōu)良種質(zhì)資源面臨稀缺。同時(shí),針對(duì)半夏的分子遺傳轉(zhuǎn)化方面的研究還比較少,分子育種較為欠缺。為了獲得兼具藥用成分高、抗逆性和抗病性強(qiáng)的半夏新品種,本研究以荊半夏為材料,利用半夏一體成苗技術(shù)獲得大量無(wú)菌苗;然后通過(guò)優(yōu)化的農(nóng)桿菌介導(dǎo)轉(zhuǎn)化體系,將來(lái)源于擬南芥(Arabidopsis thaliana)的抗熱蛋白ERECTA(ER)、來(lái)源于金黃色葡萄球菌(Staphylococcus aureus)有助于提高生物堿表達(dá)量的高活性3-羥基-3-甲基戊二酰還原輔酶A(3-hydroxy-3-methylglutaryl-Co Areduct-as e,HMGR)、來(lái)源于甜菜夜蛾(Spodoptera exigua)有助于提高植物抗逆能力的凋亡抑制因子(Inhibitor of apoptosis proteins,IAPs)、抗除草劑雙丙氨酰磷抗性蛋白(Bioiaphos resistance,BAR)等編碼基因轉(zhuǎn)化到半夏植株中,獲得基因組整合的半夏植株。同時(shí)構(gòu)建HMGR和ER基因雙價(jià)高表達(dá)載體,讓這兩個(gè)基因在同一株半夏中有較高的表達(dá)。通過(guò)農(nóng)桿菌轉(zhuǎn)化后上述轉(zhuǎn)基因體系共得到301株抗性苗,提取基因組DNA,PCR驗(yàn)證后得到72株陽(yáng)性苗隨后利用實(shí)時(shí)熒光定量PCR(Real Time PCR)技術(shù)檢測(cè)基因在半夏中的表達(dá)量,發(fā)現(xiàn)轉(zhuǎn)基因半夏中幾種基因的表達(dá)量均有了較大的提高,證明上述基因均在半夏中呈高水平表達(dá)。
[Abstract]:Pinellia ternata (Thunb. Breititi) is a traditional and precious traditional Chinese medicinal material, belonging to Pinellia ternata, a genus of Pinellia ternata. The tuber of Pinellia ternata is a good medicine for relieving phlegm and dampness, relieving pain and swelling, and reducing vomiting and adverse effects. Pinellia ternata is widely distributed all over the country, but due to over-exploitation and lack of scientific and standardized planting technology, the excellent germplasm resources of Pinellia ternata are scarce. At the same time, the research on molecular genetic transformation of Pinellia ternata is less, and molecular breeding is less. In order to obtain a new variety of Pinellia ternata with high medicinal composition, high resistance to stress and disease resistance, a large number of sterile seedlings were obtained by using Pinellia ternata as a material, and then the transformation system was optimized by Agrobacterium tumefaciens. The high activity 3-hydroxy-3-methyl-glutaryl reductive coenzyme A(3-hydroxy-3-methylglutaryl-Co Areduct-as ehmgra from Arabidopsis thaliana and Staphylococcus aureus from Staphylococcus aureus, which is derived from Spodoptera exigua, is helpful. The inhibitor of apoptosis proteinsi, a inhibitor of plant stress resistance, and Bioiaphos resistance1, a herbicide resistant protein, were transformed into Pinellia ternata plants. Genomic integration of Pinellia ternata was obtained. The bivalent expression vector of HMGR and ER gene was constructed at the same time, and the two genes were highly expressed in the same strain of Pinellia ternata. A total of 301 resistant seedlings were obtained by Agrobacterium tumefaciens transformation, and 72 positive seedlings were obtained after genomic DNA-PCR verification. The expression of the gene in Pinellia ternata was detected by real-time fluorescence quantitative PCR(Real Time. It was found that the expression of several genes in the transgenic Pinellia ternata was greatly improved, which proved that the above genes were highly expressed in Pinellia ternata.
【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
【分類號(hào)】：Q943.2;S567.239

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本文編號(hào)：1905307