本文選題：外生菌根真菌 + 鋁毒��； 參考：《西南大學(xué)》2017年碩士論文

【摘要】：磷是森林植被生長所必需的大量元素之一,其需求量僅次于氮。而土壤酸化引起的活性鋁數(shù)量增多是土壤有效磷含量降低的主要原因之一。土壤有效磷的缺乏,嚴(yán)重限制了植物的正常生長。優(yōu)良的外生菌根真菌(ECMF)可與大多數(shù)林木形成共生體,在提高植物抗鋁毒能力,促進(jìn)植物對養(yǎng)分元素特別是磷的吸收方面表現(xiàn)突出。試驗選取了3種不同的ECMF——彩色豆馬勃(Pisolithus tinctorius 715,簡稱Pt715)、松乳菇的兩個株系(Lactarius deliciosus 2,簡稱Ld2;Lactarius deliciosus 3,簡稱Ld3)為研究對象,在液體純培養(yǎng)條件下,研究了酸鋁脅迫下難溶性磷對ECMF的生長、氮磷鋁吸收、無機(jī)磷形態(tài)、抗氧化酶活性的影響,以及進(jìn)行了菌株吸收鋁、磷的動力學(xué)研究,以篩選出對難溶性磷利用效率高的優(yōu)良ECMF,同時探索ECMF對難溶性磷的活化作用及機(jī)理,為我國酸性富鋁林地上的植被修復(fù)提供理論和實踐依據(jù)。具體的研究結(jié)果如下:(1)3種供試菌株均能耐受酸鋁脅迫,且Fe-P處理對菌絲體生長的促進(jìn)效果最明顯。其中,Ld2表現(xiàn)出較強(qiáng)的耐鋁性,在P0、Na-P、Fe-P和Al-P 4種處理下菌絲生物量均顯著增加(P0.05),且增幅分別為Fe-P(22.31%)P0(15.87%)Na-P(8.87%)Al-P(6.91%);Pt715的菌絲生物量在Na-P、Fe-P、Al-P處理下顯著增加,且Fe-P(32.84%)Na-P(15.35%)Al-P(7.66%);Ld3的菌絲生物量在Na-P和Fe-P處理下顯著增加,且Fe-P(17.19%)Na-P(14.06%)。鋁脅迫時,難溶性Fe-P、Al-P、Na-P處理下的菌株生長均好于P0處理,且Fe-P效果最好,Al-P次之,Na-P最小。3種菌株中,生物量高低依次為Ld2Ld3Pt715,即Ld2在酸鋁脅迫下各種磷處理中的生長速度均優(yōu)于Ld3和Pt715。(2)3種菌株使培養(yǎng)液的pH值降低,但鋁脅迫下pH升高。鋁脅迫時,Fe-P和Al-P處理下的菌液pH值均小于Na-P,推測ECMF可能通過分泌H+或有機(jī)酸溶解難溶性Fe-P和Al-P、絡(luò)合Al3+、調(diào)節(jié)培養(yǎng)環(huán)境的pH值來達(dá)到抗鋁解磷作用。(3)Fe-P和Al-P均促進(jìn)菌絲體對鋁、總磷、Fe-P、Al-P的吸收,但抑制對氮的吸收。酸鋁脅迫下菌株對氮、鋁、總磷及Al-P的吸收增加。無鋁時,3種菌株在Fe-P處理下菌絲體中Fe-P含量最高,但鋁脅迫卻使其顯著降低,而且各菌株在4種磷源處理下均為Al-P含量及占比最高,推測鋁可能競爭鐵的吸附位點且與磷結(jié)合有優(yōu)先選擇性,進(jìn)而使Fe-P化合物或其他磷形態(tài)含量降低。3種菌株中,菌絲體內(nèi)含磷量高低依次為Ld3Ld2Pt715,即Ld3對磷的利用能力強(qiáng)于Ld2和Pt715。(4)酸鋁脅迫顯著提高ECMF的SOD、CAT酶活性,但降低POD酶活性(P0.05)。難溶性Fe-P、Al-P處理下的菌絲體SOD、POD、CAT酶活性均高于Na-P,說明難溶性磷處理有利于抗氧化酶升高;鋁脅迫使Ld2的SOD、CAT酶活性增幅均大于Ld3和Pt715,POD酶活性降幅均小于Ld3和Pt715,推測Ld2的抗鋁脅迫能力較強(qiáng)。(5)相關(guān)性分析表明,3種ECMF的總P、Al、Al-P含量呈兩兩極顯著正相關(guān)(P0.01),說明P與Al的吸收有協(xié)同促進(jìn)作用,Al-P隨總P和Al含量的增多而增多,推測Al和P在進(jìn)入菌絲體內(nèi)仍可能相互結(jié)合。SOD與CAT酶活性呈極顯著正相關(guān),說明SOD與CAT協(xié)同促進(jìn)消除抗氧自由基。SOD、CAT酶活性與P、Al-P含量呈顯著正相關(guān)(P0.05),說明菌株通過提高SOD、CAT酶活性,抗鋁脅迫和活化難溶性磷能力增強(qiáng)。(6)3種菌株的CEC大小依次為Ld3≥Ld2Pt715,推測Ld3和Ld2對Al3+的吸附量大于Pt715�？偟膩砜�,Ld3、Ld2吸收鋁的Km大、Imax大,表明松乳菇更能適應(yīng)高濃度鋁的土壤環(huán)境。(7)鋁脅迫下,除Ld2的C_(min)無明顯變化外,ECMF吸收磷的Km和C_(min)值均減小,Imax值均增大,即鋁增強(qiáng)了菌株吸收磷的能力,并且供試菌株均能夠適應(yīng)低磷的土壤環(huán)境。松乳菇的兩個株系Ld2和Ld3吸收磷的Km和Imax均小于Pt715,說明松乳菇更能適合低磷的土壤環(huán)境。綜上,試驗所用的3種菌株在酸鋁脅迫下均能適應(yīng)高鋁低磷環(huán)境,活化利用難溶性磷,且Ld2和Ld3的適應(yīng)能力和活化利用能力均強(qiáng)于Pt715。其中,Ld2生長速度最快、抗鋁性最強(qiáng),Ld3吸收磷的能力最強(qiáng)。3種菌主要通過分泌H+或有機(jī)酸、促進(jìn)Al-P、Fe-P溶解、增加對養(yǎng)分磷的吸收,增加對Al的吸收或吸附,絡(luò)合Al3+,提高SOD、POD、CAT酶活性,來活化酸性鋁脅迫下的難溶性磷。
[Abstract]:Phosphorus is one of the most important elements necessary for the growth of forest vegetation, and its demand is second only to nitrogen. The increase in the amount of active aluminum caused by soil acidification is one of the main reasons for reducing the content of effective phosphorus in the soil. The lack of available phosphorus seriously restricts the normal growth of plants. The excellent ectomycorrhizal fungi (ECMF) can be formed with most trees. Symbionts, in improving plant resistance to aluminum toxicity and promoting plant uptake of nutrient elements especially phosphorus, have selected 3 different kinds of ECMF - color bean (Pisolithus tinctorius 715, Pt715), two strains of Lactobacillus Pleurotus (Lactarius deliciosus 2, Ld2; Lactarius deliciosus 3, Ld3) as research Under the condition of liquid pure culture, the effects of soluble phosphorus on the growth of ECMF, the absorption of nitrogen, phosphorus and aluminum, the effect of the inorganic phosphorus form and the activity of antioxidant enzymes were studied, and the kinetics of the absorption of aluminum and phosphorus by strains was carried out to screen out the excellent ECMF with high efficiency of soluble phosphorus, and the activity of ECMF to the insoluble phosphorus was also explored. The effect and mechanism of the study provide theoretical and practical basis for the vegetation restoration on the acid rich aluminum forest in China. The results are as follows: (1) 3 strains of strain can tolerate aluminum stress, and the effect of Fe-P treatment on mycelium growth is the most obvious. Among them, Ld2 shows a strong resistance to aluminum, and 4 kinds of bacteria are treated in P0, Na-P, Fe-P and Al-P. The biomass increased significantly (P0.05), and the increase was Fe-P (22.31%) P0 (15.87%) Na-P (8.87%) Al-P (6.91%), and the mycelial biomass of Pt715 increased significantly under Na-P, Fe-P, Al-P, and Fe-P (32.84%) Na-P (15.35%) Al-P (7.66%). The growth of strain Fe-P, Al-P and Na-P was better than P0 treatment, and the effect of Fe-P was the best, Al-P was the best. In Na-P smallest.3 strain, the biomass of the strain was in turn Ld2Ld3Pt715, that is, the growth rate of Ld2 in all kinds of phosphorus treatments under aluminum acid stress was superior to Ld3 and Pt715. (2) 3 strains to reduce the value of the culture solution, but increased under aluminum stress. When aluminum stress was stressed, the pH value of the bacterial solution under Fe-P and Al-P treatment was less than Na-P. It is suggested that ECMF may dissolve difficult Fe-P and Al-P by secreting H+ or organic acid, complexing Al3+ and adjusting the pH value of the culture environment to achieve the effect of anti aluminum phosphate hydrolysis. (3) Fe-P and Al-P all promote the absorption of aluminum, total phosphorus, and nitrogen, but inhibit the absorption of nitrogen. Aluminum stress The absorption of nitrogen, aluminum, total phosphorus and Al-P by the lower strains increased. When the 3 strains were free of aluminum, the content of Fe-P was the highest in the hypha under Fe-P treatment, but the aluminum stress was significantly reduced, and all the strains were Al-P content and the highest proportion under the treatment of 4 kinds of phosphorus sources. Among the Fe-P compounds or other phosphorus forms, the level of phosphorus content in the hyphae of.3 strains was Ld3Ld2Pt715, that is, the utilization of Ld3 to phosphorus is stronger than that of Ld2 and Pt715. (4). The activity of SOD and CAT enzyme in ECMF is significantly increased, but the POD enzyme activity (P0.05) is reduced. Na-P, indicating that the treatment of soluble phosphorus is beneficial to the increase of antioxidant enzymes; aluminum coerced Ld2 SOD, CAT enzyme activity increased more than Ld3 and Pt715, POD enzyme activity decreased less than Ld3 and Pt715, and speculates that Ld2 is stronger in resistance to aluminum stress. (5) correlation analysis shows that the total P of the 3 ECMF is two and two poles. Al-P increased with the increase of total P and Al content. It is suggested that Al and P may still have a significant positive correlation with the activity of.SOD and CAT enzyme in the hyphae into the mycelium, indicating that SOD and CAT synergistically promote the elimination of oxygen free radicals,.SOD, and the activity of CAT enzyme is significantly positively correlated with P. The activity of AT enzyme, resistance to aluminum stress and activation of insoluble phosphorus increased. (6) the CEC size of 3 strains was Ld3 > Ld2Pt715 in turn. The adsorption amount of Ld3 and Ld2 on Al3+ was greater than Pt715., Ld3, Ld2 absorption of aluminum was large and Imax was large, indicating that the mushroom could adapt to the soil environment of high concentration aluminum. (7) there was no obvious change under aluminum stress. In addition, the Km and C_ (min) value of ECMF absorption decreased and the Imax value increased, that is, aluminum enhanced the ability to absorb phosphorus, and the tested strains were able to adapt to the low phosphorus soil environment. The Km and Imax of the two strains of Lactobacillus Pleurotus, Ld2 and Ld3, were less than Pt715, indicating that Lactobacillus Pleurotus can be more suitable for low phosphorus soil environment. The 3 strains can adapt to the environment of high aluminum and low phosphorus under the stress of aluminum and activate the refractory phosphorus, and the adaptability and activation ability of Ld2 and Ld3 are stronger than that of Pt715.. Ld2 has the fastest growth rate and the strongest aluminum resistance. The strongest.3 bacteria of Ld3 absorb the H+ or organic acids, which can promote Al-P, Fe-P dissolve and increase the nutrition. The absorption of phosphorus increased the absorption and adsorption of Al, complexed Al3+, enhanced SOD, POD and CAT enzyme activities to activate the insoluble phosphorus under acidic aluminum stress.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S718.81

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