本文選題：雷公藤 + 宿主生長��； 參考：《福建農(nóng)林大學(xué)》2017年碩士論文

【摘要】：雷公藤(Tripterygium wilfordii)是珍貴的傳統(tǒng)藥用植物之一,體內(nèi)有多種活性物質(zhì),具有抗炎、抗菌、抗艾滋病毒、抗腫瘤和免疫抑制等作用。然而由于野生雷公藤資源蘊(yùn)藏量少,加之人工栽培生長期長,該藥用植物已遠(yuǎn)遠(yuǎn)無法滿足國內(nèi)外市場的需求。相關(guān)研究表明,內(nèi)生菌對(duì)宿主植物的生理生長產(chǎn)生重要影響,促進(jìn)鐵載體的合成、分泌吲哚乙酸和溶磷的產(chǎn)生,增強(qiáng)宿主植物的抗性,參與合成宿主植物相同或相近的次生代謝產(chǎn)物。本研究通過對(duì)雷公藤內(nèi)生細(xì)菌與內(nèi)生真菌及其組合有益生物活性進(jìn)行評(píng)價(jià),以及與宿主組培苗共生體系的建立和細(xì)胞懸浮誘導(dǎo)子添加技術(shù),研究雷公藤內(nèi)生真菌和內(nèi)生細(xì)菌組合對(duì)宿主植株和細(xì)胞生長、生理生化特征以及活性物質(zhì)合成的影響。主要研究結(jié)果如下:(1)對(duì)雷公藤內(nèi)生真菌Fusariuum oxysporum NS33和Penicilliurr steckii NS6、內(nèi)生細(xì)菌 Enterobacter cloacae sub sp LG3 和 Serratia marcescensLY1以及真菌與細(xì)菌的四個(gè)組合NS33-LG3、NS33-LY1、NS6-LG3、NS6-LY1的促生特性及促進(jìn)小麥萌發(fā)的作用進(jìn)行了評(píng)價(jià),內(nèi)生真菌NS6、NS33及混合菌株NS33-LG3具有較高的產(chǎn)鐵載體能力和IAA分泌能力;混合菌株NS33-LY1具有較高的產(chǎn)鐵載體能力和溶磷能力;而菌株LY1亦顯示出較高的溶磷能力。多數(shù)菌株及菌株組合顯著促進(jìn)了小麥種子萌發(fā)、芽伸長、根系活力及幼苗葉綠素含量;菌株NS33與菌株LG3能夠協(xié)同促進(jìn)IAA的分泌,兩者混合接種的小麥幼苗根系活力得到進(jìn)一步提高。(2)將目標(biāo)菌株及組合與雷公藤細(xì)胞分別進(jìn)行懸浮共培養(yǎng),研究了不同共培養(yǎng)體系雷公藤細(xì)胞的生長及其生理生化特征。在共培養(yǎng)前期,與對(duì)照相比,接種單一內(nèi)生菌株提高了細(xì)胞的干重,其中菌株NS6的促生效果最明顯;而在共培養(yǎng)后期,無論是單一內(nèi)生菌還是混合內(nèi)生菌均對(duì)細(xì)胞生長具有抑制作用。內(nèi)生真菌和菌株組合處理的培養(yǎng)液pH值有明顯的升高,而內(nèi)生細(xì)菌LY1則明顯降低了培養(yǎng)液的pH值,其具有產(chǎn)酸性。另外,當(dāng)雷公藤細(xì)胞同混合菌株共培養(yǎng)時(shí),培養(yǎng)基中總糖消耗量是最大的,而接種單獨(dú)菌株時(shí)則對(duì)細(xì)胞可溶性蛋白含量具有一定的提高作用。接種內(nèi)生菌會(huì)影響雷公藤細(xì)胞的POD、CAT及SOD活性,與對(duì)照相比,接種單獨(dú)菌株會(huì)更加提升POD與CAT的活性,而細(xì)胞MDA含量則明顯下降。(3)將目標(biāo)菌株及其組合制備成菌液(JY)、菌體(JT)誘導(dǎo)子添加至雷公藤細(xì)胞懸浮培養(yǎng)體系中,各個(gè)誘導(dǎo)子對(duì)懸浮細(xì)胞的鮮重具有促進(jìn)作用,并且降低了培養(yǎng)液的pH,其中比較顯著地是LG3-JY、LY1-JY誘導(dǎo)子。JY誘導(dǎo)子提高了培養(yǎng)液電導(dǎo)率,而JT誘導(dǎo)子降低了培養(yǎng)液電導(dǎo)率。各誘導(dǎo)子均能提高懸浮細(xì)胞培養(yǎng)液中營養(yǎng)成分的消耗速率。在不同培養(yǎng)階段,誘導(dǎo)子添加對(duì)細(xì)胞POD、CAT、SOD活性的影響不同,而MDA含量則呈有規(guī)律的先增加后降低的變化。(4)將目標(biāo)菌株及其組合接種于雷公藤組培苗,8種接種處理對(duì)組培苗的生長均具有促進(jìn)作用。內(nèi)生真菌比內(nèi)生細(xì)菌的促進(jìn)作用更強(qiáng),多數(shù)內(nèi)生真菌和內(nèi)生細(xì)菌的混合接種相對(duì)于單獨(dú)接種對(duì)雷公藤組培苗生長的促進(jìn)效應(yīng)沒有明顯的提高或降低。接種內(nèi)生菌能明顯提高雷公藤組培苗的葉綠素和可溶性蛋白含量,然而卻降低了體內(nèi)可溶性糖含量。接種內(nèi)生菌及其組合降低了組培苗CAT、MDA、超氧陰離子自由基活力,混合菌株的降低作用更明顯。(5)接種目標(biāo)菌株及其組合的雷公藤組培苗對(duì)C、N、P、K營養(yǎng)元素的吸收發(fā)生了不同程度的變化。接種各菌株及菌株組合的組培苗體內(nèi)C、K含量明顯提高,而接種內(nèi)生細(xì)菌LG3、LY1促進(jìn)了組培苗N積累,接種菌株NS33、LG3、LY1及菌株組合NS33-LY1有利于組培苗P含量的增加。相對(duì)于單獨(dú)接種,混合接種內(nèi)生細(xì)菌和內(nèi)生真菌對(duì)雷公藤組培苗營養(yǎng)元素的吸收即有正效應(yīng)亦有負(fù)效應(yīng)。各菌株及其組合均能明顯提高雷公藤組培苗中甲素和紅素的積累,其中混合菌株NS33-LG3和菌株LG3的作用最明顯�；旌辖臃N菌株NS33和LG3、NS6和LY1對(duì)雷公藤紅素的積累表現(xiàn)出正效應(yīng)。
[Abstract]:Lei Gongteng (Tripterygium wilfordii) is one of the precious traditional medicinal plants. There are many active substances in the body, which have anti - inflammatory, anti - bacterial, anti - HIV, anti-tumor and immunosuppressive effects. However, because of the few reserves of wild Lei Gongteng resources and long growing period of artificial cultivation, the medicinal plant has been far from meeting the domestic and foreign markets. Requirements. Related studies have shown that endophytes have important effects on the physiological growth of host plants, promote the synthesis of iron carriers, secrete the production of indoliacetic acid and phosphorus, enhance the resistance of the host plants and participate in the same or similar secondary metabolites in the synthetic host plants. This study has been conducted through the study of endophytic and endophytic fungi and their groups in the study of Tripterygium wilfordii. The effects of beneficial biological activity, the establishment of the symbiotic system with the host tissue culture and the addition of cell suspension inducers were used to study the effects of the endophytic and endophytic bacteria of Tripterygium wilfordii on the plant and cell growth, physiological and biochemical characteristics and the synthesis of active substances. The main results are as follows: (1) the endophytic fungi of Tripterygium wilfordii Fusariuum oxysporum NS33 and Penicilliurr steckii NS6, endogenous bacteria Enterobacter cloacae sub sp LG3 and Serratia, as well as the four combinations of fungi and bacteria. The hybrid strain NS33-LY1 had a high ability to produce iron and the ability to dissolve the phosphorus, and the strain LY1 also showed higher ability to dissolve the phosphorus, while the strain and strain combination significantly promoted the germination of the wheat seed, the elongation of the bud, the vigor of the root and the chlorophyll content of the seedling, and the strain NS33 and the strain LG3 of the strain of strain LY1. The root activity of wheat seedlings with mixed inoculation was further improved. (2) the growth and physiological and biochemical characteristics of Tripterygium wilfordii cells in different co culture systems were studied by suspension co culture of the target strains and combinations and Tripterygium wilfordii cells. In the pre culture period, the inoculation was compared with the control. The bacterial strain increased the dry weight of the cell, of which the strain NS6 was most effective in promoting the growth, while in the late stage of co culture, both endophyte and mixed endophyte had inhibitory effect on the cell growth. The pH value of the endophyte and strain combination treatment was significantly higher, while the endophyte LY1 significantly reduced the pH of the culture solution. In addition, when the Tripterygium wilfordii cells were co cultured with the mixed strain, the total sugar consumption was the largest in the medium, while the inoculation of isolated strains had a certain effect on the soluble protein content of the cells. The inoculation of endophyte could affect the activity of POD, CAT and SOD of Tripterygium wilfordii cells, and inoculated the isolated strains compared with the control. The activity of POD and CAT would be enhanced and the MDA content of cells decreased significantly. (3) the target strains and their combinations were prepared to produce bacterial liquid (JY), and the JT inducer was added to the suspension culture system of Tripterygium wilfordii cells. Each inducer could promote the fresh weight of the suspension cells and reduced the pH of the culture medium, which was more notably LG3-. JY, the inducer of the LY1-JY inducer increased the conductivity of the culture medium, while the JT inducer reduced the conductivity of the culture medium. All the inducers could increase the consumption rate of nutrient components in the suspension cell culture medium. The effects of the inducers on the activity of POD, CAT and SOD in the cells were different at different stages of culture, while the MDA content increased first. (4) the target strains and their combinations were inoculated to the tissue culture seedlings of Tripterygium wilfordii, and the 8 inoculation treatments could promote the growth of the tissue culture seedlings. The endophytic fungi were more effective than endophytic bacteria, and the mixed inoculation of most endophyte and endophytic bacteria had no effect on the growth of the plantlets of Tripterygium wilfordii. The inoculation of endophyte could significantly increase the content of chlorophyll and soluble protein in the tissue culture of Tripterygium wilfordii, but decreased the content of soluble sugar in the body. The inoculation of endophyte and its combination reduced the CAT, MDA, superoxide anion free radical activity of the tissue culture, and the effect of the mixed strain was more obvious. (5) the inoculated target strains and The combination of Tripterygium wilfordii tissue culture seedlings had different changes in the absorption of C, N, P, K nutrient elements. The content of C and K in the tissue culture seedlings of each strain and strain combination increased obviously, while the inoculated endophyte LG3, LY1 promoted the N accumulation of the tissue culture seedlings, the inoculated strains NS33, LG3, LY1 and strain combinations were beneficial to the increase of the content of tissue culture seedlings. Compared with the individual inoculation, the Mixed Inoculation endophyte and endophyte had positive and negative effects on the nutrient absorption of the Tripterygium wilfordii tissue culture seedlings. All strains and their combinations could obviously improve the accumulation of methyl and erythropoietin in the tissue culture of Tripterygium wilfordii, of which the mixed strain NS33-LG3 and the strain LG3 were most obvious. The Mixed Inoculation strain NS33 And LG3, NS6 and LY1 showed positive effects on the accumulation of Tripterygium.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S567.19

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