内生细菌与内生真菌组合对雷公藤生长及生理生化特性的影响

发布时间：2018-07-09 09:25

本文选题：雷公藤 + 宿主生长　；参考：《福建农林大学》2017年硕士论文

【摘要】：雷公藤(Tripterygium wilfordii)是珍贵的传统药用植物之一,体内有多种活性物质,具有抗炎、抗菌、抗艾滋病毒、抗肿瘤和免疫抑制等作用。然而由于野生雷公藤资源蕴藏量少,加之人工栽培生长期长,该药用植物已远远无法满足国内外市场的需求。相关研究表明,内生菌对宿主植物的生理生长产生重要影响,促进铁载体的合成、分泌吲哚乙酸和溶磷的产生,增强宿主植物的抗性,参与合成宿主植物相同或相近的次生代谢产物。本研究通过对雷公藤内生细菌与内生真菌及其组合有益生物活性进行评价,以及与宿主组培苗共生体系的建立和细胞悬浮诱导子添加技术,研究雷公藤内生真菌和内生细菌组合对宿主植株和细胞生长、生理生化特征以及活性物质合成的影响。主要研究结果如下:(1)对雷公藤内生真菌Fusariuum oxysporum NS33和Penicilliurr steckii NS6、内生细菌 Enterobacter cloacae sub sp LG3 和 Serratia marcescensLY1以及真菌与细菌的四个组合NS33-LG3、NS33-LY1、NS6-LG3、NS6-LY1的促生特性及促进小麦萌发的作用进行了评价,内生真菌NS6、NS33及混合菌株NS33-LG3具有较高的产铁载体能力和IAA分泌能力;混合菌株NS33-LY1具有较高的产铁载体能力和溶磷能力;而菌株LY1亦显示出较高的溶磷能力。多数菌株及菌株组合显著促进了小麦种子萌发、芽伸长、根系活力及幼苗叶绿素含量;菌株NS33与菌株LG3能够协同促进IAA的分泌,两者混合接种的小麦幼苗根系活力得到进一步提高。(2)将目标菌株及组合与雷公藤细胞分别进行悬浮共培养,研究了不同共培养体系雷公藤细胞的生长及其生理生化特征。在共培养前期,与对照相比,接种单一内生菌株提高了细胞的干重,其中菌株NS6的促生效果最明显;而在共培养后期,无论是单一内生菌还是混合内生菌均对细胞生长具有抑制作用。内生真菌和菌株组合处理的培养液pH值有明显的升高,而内生细菌LY1则明显降低了培养液的pH值,其具有产酸性。另外,当雷公藤细胞同混合菌株共培养时,培养基中总糖消耗量是最大的,而接种单独菌株时则对细胞可溶性蛋白含量具有一定的提高作用。接种内生菌会影响雷公藤细胞的POD、CAT及SOD活性,与对照相比,接种单独菌株会更加提升POD与CAT的活性,而细胞MDA含量则明显下降。(3)将目标菌株及其组合制备成菌液(JY)、菌体(JT)诱导子添加至雷公藤细胞悬浮培养体系中,各个诱导子对悬浮细胞的鲜重具有促进作用,并且降低了培养液的pH,其中比较显著地是LG3-JY、LY1-JY诱导子。JY诱导子提高了培养液电导率,而JT诱导子降低了培养液电导率。各诱导子均能提高悬浮细胞培养液中营养成分的消耗速率。在不同培养阶段,诱导子添加对细胞POD、CAT、SOD活性的影响不同,而MDA含量则呈有规律的先增加后降低的变化。(4)将目标菌株及其组合接种于雷公藤组培苗,8种接种处理对组培苗的生长均具有促进作用。内生真菌比内生细菌的促进作用更强,多数内生真菌和内生细菌的混合接种相对于单独接种对雷公藤组培苗生长的促进效应没有明显的提高或降低。接种内生菌能明显提高雷公藤组培苗的叶绿素和可溶性蛋白含量,然而却降低了体内可溶性糖含量。接种内生菌及其组合降低了组培苗CAT、MDA、超氧阴离子自由基活力,混合菌株的降低作用更明显。(5)接种目标菌株及其组合的雷公藤组培苗对C、N、P、K营养元素的吸收发生了不同程度的变化。接种各菌株及菌株组合的组培苗体内C、K含量明显提高,而接种内生细菌LG3、LY1促进了组培苗N积累,接种菌株NS33、LG3、LY1及菌株组合NS33-LY1有利于组培苗P含量的增加。相对于单独接种,混合接种内生细菌和内生真菌对雷公藤组培苗营养元素的吸收即有正效应亦有负效应。各菌株及其组合均能明显提高雷公藤组培苗中甲素和红素的积累,其中混合菌株NS33-LG3和菌株LG3的作用最明显。混合接种菌株NS33和LG3、NS6和LY1对雷公藤红素的积累表现出正效应。
[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.
【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567.19

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