發(fā)布時(shí)間：2018-08-15 16:34

【摘要】：磷(phosphorus,P)是自然生態(tài)系統(tǒng)和農(nóng)業(yè)生態(tài)系統(tǒng)中一種非常重要的大量元素。全國(guó)土壤普查統(tǒng)計(jì),約有2/3的土壤依舊缺磷,土壤缺磷成為限制作物高產(chǎn)的主要影響因素。隨著糧食危機(jī)及環(huán)境危機(jī)的日益加劇,如何在保護(hù)環(huán)境的基礎(chǔ)上提高農(nóng)作物產(chǎn)量已成為全球乃至全人類面臨的一個(gè)主要問(wèn)題。解磷細(xì)菌(phosphate-solubilizing bacteria,PSB)是人們較早發(fā)現(xiàn)并被廣泛利用的一類植物促生菌(plant growth-promoting bacteria,PGPB),能促進(jìn)土壤難溶性磷的溶解,提高土壤磷素的利用效率,對(duì)環(huán)境危害極小,在促進(jìn)作物生長(zhǎng)方面具有極好的效果。本試驗(yàn)選用實(shí)驗(yàn)室前期分離的一株有機(jī)無(wú)機(jī)雙解解磷細(xì)菌WY4,經(jīng)16s rRNA鑒定為巨大芽孢桿菌(Bacillus megaterium),利用綠色熒光蛋白標(biāo)記(green fluorescent protein,GFP)技術(shù)對(duì)WY4進(jìn)行熒光標(biāo)記,以期研究解磷細(xì)菌在小白菜根系及根系土壤中的定殖動(dòng)態(tài),同時(shí)研究解磷細(xì)菌WY4在大田條件下對(duì)小白菜(Brassica rapa L.Chinensis Group.)生長(zhǎng)、品質(zhì)及土壤養(yǎng)分的影響。主要研究結(jié)果如下:1.綠色熒光蛋白標(biāo)記解磷細(xì)菌WY4,獲得具有較為穩(wěn)定遺傳性的標(biāo)記菌株WY4-GFP。標(biāo)記菌株WY4-GFP與野生型菌株WY4在生長(zhǎng)動(dòng)態(tài)、有機(jī)酸分泌及解磷活性均無(wú)顯著性差異,說(shuō)明GFP標(biāo)記不影響WY4菌株的生理活性,可以替代野生型菌株。2.WY4-GFP在小白菜根際土壤及根系組織中均可定殖。(1)在小白菜根際土中接種30 d后,WY4-GFP定殖數(shù)量為5.80×104 CFU/g,說(shuō)明WY4-GFP在小白菜根際土壤中具有持久穩(wěn)定的定殖能力;(2)WY4-GFP在小白菜根冠及分生區(qū)大量定殖,在伸長(zhǎng)區(qū)及側(cè)根根毛處數(shù)量較少,同時(shí)根表皮細(xì)胞間隙上也有較多的標(biāo)記菌株,表明WY4-GFP在小白菜根系組織中具有良好的定殖能力;試驗(yàn)結(jié)果同時(shí)表明WY4-GFP與WY4均能促進(jìn)小白菜的生長(zhǎng),且二者之間無(wú)顯著差異。這為后期深入研究解磷細(xì)菌在土壤中的定殖奠定了基礎(chǔ)。3.大田條件下,解磷細(xì)菌WY4仍具有較好的解磷能力。施用解磷細(xì)菌WY4后顯著提升土壤速效磷含量,降低土壤pH,且土壤速效鉀、有機(jī)質(zhì)、硝態(tài)氮、銨態(tài)氮含量也顯著提高;小白菜根長(zhǎng)、株高、葉片數(shù)、生物量與對(duì)照差異顯著增加;小白菜葉片硝態(tài)氮含量顯著降低,可溶性糖、維生素C、纖維素等含量顯著增加,小白菜品質(zhì)明顯改善。表明解磷細(xì)菌WY4能夠顯著提高土壤磷素利用效率,改善土壤品質(zhì),促進(jìn)小白菜生長(zhǎng)。
[Abstract]:Phosphorus (P) is a very important element in natural and agricultural ecosystems. According to the national soil census, about two thirds of the soils are still deficient in phosphorus, and phosphorus deficiency is the main factor limiting the high yield of crops. With the increasing of food crisis and environmental crisis, how to improve crop yield on the basis of environmental protection has become a major problem facing the whole world and all mankind. Phosphate-solubilizing bacterium PSB is a kind of plant promoting bacteria (plant growth-promoting bacterium PGPB), which has been discovered and widely used by people earlier. It can promote the dissolution of insoluble phosphorus in soil, improve the utilization efficiency of soil phosphorus, and do little harm to the environment. It has excellent effect in promoting crop growth. In this experiment, a strain of organic-inorganic biolysis bacteria WY4 was isolated in the laboratory and identified as Bacillus megaterium (Bacillus megaterium), by using green fluorescent protein labeling (GFP) technique to label WY4 by 16s rRNA. The purpose of this study was to study the colonization dynamics of phosphorus releasing bacteria in Chinese cabbage root system and root system soil, and to study the effect of phosphorus releasing bacteria WY4 on (Brassica rapa L.Chinensis group of Chinese cabbage under field conditions. Effects of growth, quality and soil nutrients. The main results are as follows: 1. A stable genetic marker strain WY4-GFPwas obtained by labeling phosphobacteria WY4 with green fluorescent protein. There was no significant difference in growth dynamics, organic acid secretion and phosphorus-releasing activity between the labeled strain WY4-GFP and wild-type strain WY4, indicating that GFP labeling did not affect the physiological activity of WY4 strain. WY4-GFP can be substituted for wild-type strain .2.WY4-GFP can colonize in rhizosphere soil and root tissue of pakchoi. (1) after inoculating in rhizosphere soil of pakchoi for 30 days, the colonization quantity of WY4-GFP is 5.80 脳 10 ~ 4 CFU / g, which indicates that WY4-GFP has lasting and stable colonization in rhizosphere soil of pakchoi. (2) colonization of WY4-GFP in root crown and meristem of pakchoi, At the same time, there were more labeled strains in the intercellular space of root epidermis, which indicated that WY4-GFP had a good colonization ability in the root tissue of pakchoi (Brassica campestris). The results also showed that both WY4-GFP and WY4 could promote the growth of Chinese cabbage, and there was no significant difference between them. This laid a foundation for further study on the colonization of phosphorus releasing bacteria in soil. Under the field conditions, the phosphorus releasing bacteria (WY4) still had good phosphorus releasing ability. The content of available phosphorus, soil pH, available potassium, organic matter, nitrate nitrogen and ammonium nitrogen in soil were significantly increased after WY4 application, and the root length, plant height, leaf number, biomass and biomass of pakchoi increased significantly. The contents of nitrate nitrogen, soluble sugar, vitamin C and cellulose in leaves of Chinese cabbage were significantly decreased, and the quality of Chinese cabbage was improved obviously. The results showed that WY4 could significantly improve the utilization efficiency of phosphorus in soil, improve soil quality and promote the growth of pakchoi.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S182;S634.3

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