【摘要】：油料植物蓖麻(Ricinus communist L.)有著良好的適應(yīng)性、重金屬耐性及高產(chǎn)油率,可應(yīng)用于重金屬污染的植物修復(fù)。而利用蓖麻進(jìn)行修復(fù)時(shí),高濃度的重金屬離子會(huì)破壞植物的正常代謝,限制種子萌發(fā),從而對(duì)其生長(zhǎng)生存和修復(fù)效果造成嚴(yán)重影響。針對(duì)以上情況,本文以蓖麻為供試植物,并不同于以往大都采用單一微生物進(jìn)行聯(lián)合修復(fù)的傳統(tǒng)方法,構(gòu)建了復(fù)合菌群,研究了微生物菌群對(duì)重金屬脅迫下蓖麻種子萌發(fā)的影響,論文取得的主要研究結(jié)果如下:(1)涂布初篩后,共從課題組內(nèi)經(jīng)菌肥改良后的向日葵、辣椒根際土壤中分離出真菌10株,細(xì)菌24株,選取形態(tài)穩(wěn)定、長(zhǎng)勢(shì)良好的7株微生物以促生性能為指標(biāo)進(jìn)行復(fù)篩,初步確定L6、L7、Lll、L14與X85株菌株具備良好的促生性能,可作為構(gòu)建復(fù)合菌群的可選根際促生菌。(2)以耐性菌為基礎(chǔ),研究鉛鋅脅迫下復(fù)合菌群對(duì)蓖麻種子萌發(fā)的影響。對(duì)課題組前期研究的三株耐性真菌HA、WB、J3,通過(guò)正交實(shí)驗(yàn),考查其不同組合對(duì)鉛鋅的吸附性能。結(jié)果顯示,HA與J3混合培養(yǎng)(X2)在最適條件下,對(duì)鉛離子的去除率可達(dá)91.3%,對(duì)鋅離子的去除率可達(dá)51.4%,高于其他組合,能滿足復(fù)合菌群對(duì)鉛鋅脅迫的耐性需求,因而可以將HA、J3作為構(gòu)建復(fù)合功能菌群的固定菌株。根據(jù)課題組已有研究,耐性真菌HA為米曲霉(Aspergillus oryzae),耐性真菌J3為蟲(chóng)生輪殖孢(Verticillium insectorum)。(3)對(duì)篩選出的根際促生菌與耐性真菌進(jìn)行平板對(duì)峙實(shí)驗(yàn),結(jié)果表明,復(fù)篩出的5株微生物僅L6、L11與HA之間有一定的拮抗作用,結(jié)合促生性能,優(yōu)選X8、L14作為構(gòu)建符合菌群的根際促生菌,并進(jìn)行測(cè)序鑒定。根據(jù)測(cè)序結(jié)果構(gòu)建系統(tǒng)發(fā)育樹(shù),進(jìn)行BLAST比對(duì)后最終確定L14為尖孢鐮刀菌(Fusarium oxysporum),而X8 為枯草芽孢桿菌(Bacillus subtilis)。(4)通過(guò)沙培實(shí)驗(yàn),考察微生物菌群對(duì)重金屬脅迫下蓖麻種子萌發(fā)的影響,包括接種復(fù)合菌群對(duì)蓖麻生物量、重金屬富集及根系生長(zhǎng)等情況的影響,探討其對(duì)蓖麻種子萌發(fā)及幼苗生長(zhǎng)的作用并優(yōu)選出最適菌群。結(jié)果表明,復(fù)合菌群的添加可以有效促進(jìn)植物對(duì)重金屬的富集,其中,D1菌群(HA、J3、L14)在200mg/L條件下對(duì)Pb的累積量提升達(dá)到37.1%,在400mg/L條件下對(duì)Zn的累積量提升達(dá)到26.9%;D2(HA、J3、X8)菌群在600mg/L條件下對(duì)Pb的累積量提升達(dá)到29.3%,在400mg/L條件下對(duì)Zn的累積量提升達(dá)到41.6%。綜合對(duì)比復(fù)合菌群D1與D2的各項(xiàng)生長(zhǎng)指標(biāo)、根系形態(tài)及重金屬富集量,優(yōu)選出復(fù)合功能菌群D2為最優(yōu)菌群,可作為下一步盆栽實(shí)驗(yàn)的復(fù)合功能菌群。最終結(jié)果表明,該菌群可以有效促進(jìn)鉛鋅脅迫下蓖麻種子的萌發(fā)。
[Abstract]:Oil plant castor (Ricinus communist L.) It has good adaptability, heavy metal tolerance and high oil yield, and can be used in phytoremediation of heavy metal pollution. However, high concentration of heavy metal ions will destroy the normal metabolism of plants and restrict the seed germination, which will seriously affect the growth, survival and repair effect of castor. In view of the above situation, the compound flora was constructed by using castor as the tested plant, which was different from the traditional method in which single microorganism was used to repair the plant in the past. The effects of microbial flora on the germination of castor seeds under heavy metal stress were studied. The main results were as follows: (1) after primary screening, sunflower was improved by bacterial fertilizer. 10 strains of fungi and 24 strains of bacteria were isolated from rhizosphere soil of capsicum. 7 strains of microbes with stable morphology and good growth were selected and screened for promoting growth ability. It was preliminarily confirmed that L6, L7, LlllL14 and X85 strains had good growth promoting performance. (2) on the basis of tolerant bacteria, the effect of compound flora on seed germination of castor under lead and zinc stress was studied. The adsorption properties of three resistant fungi HA,WB,J3, for lead and zinc were studied by orthogonal experiment. The results showed that under the optimum conditions, the removal rate of lead ion and zinc ion in HA and J3 mixed culture (X2) was 91.3 and 51.4, which was higher than that in other combinations. Therefore, HA,J3 can be used as a fixed strain to construct complex functional flora. According to the previous research, the resistant fungus HA is (Aspergillus oryzae), resistant fungus J3 is (Verticillium insectorum). (3. The results show that the isolated rhizosphere probiotic fungi and tolerant fungi are confronted with each other. Only 5 strains of microorganism L6, L11 and HA had antagonistic effect. Combined with the growth promoting ability, X8G L14 was selected as the rhizosphere promoting bacteria which was in accordance with the bacterial flora, and identified by sequencing. The phylogenetic tree was constructed according to the sequencing results. After BLAST comparison, it was determined that L14 was (Fusarium oxysporum), and X8 was (Bacillus subtilis). (4 (Bacillus subtilis). The effects of microbial flora on the germination of castor seeds under heavy metal stress were investigated, including the effects of inoculation on ricin biomass, accumulation of heavy metals and root growth. To study the effect of castor on seed germination and seedling growth and to select the optimal flora. The results showed that the addition of complex flora could effectively promote the accumulation of heavy metals in plants. Among them, the accumulation of Pb by D1 microflora (HA,J3,L14) reached 37.1g under 200mg/L condition. Under the condition of 400mg/L, the cumulative amount of Zn increased to 26.9%. The cumulative amount of D2 (HA,J3,X8) to Pb increased to 29.3 under 600mg/L and 41.6 to Zn under 400mg/L. By comparing the growth indexes, root morphology and heavy metal accumulation of the complex bacteria D 1 and D 2, the composite functional bacteria D 2 was selected as the optimal flora, which could be used as the next step in the pot experiment. The results showed that the microflora could effectively promote the germination of castor seeds under lead and zinc stress.
【學(xué)位授予單位】：中南林業(yè)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)】：S565.6;X17

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