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Archive > Volume 16 Issue 2 > 2024,16(2):214-226. DOI:https://doi.org/10.1016/j.chmed.2022.11.004 Prev Next

中度干旱下叢枝菌根真菌和促生細(xì)菌協(xié)作促進(jìn)重樓皂苷積累的研究

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Arbuscular mycorrhiza and plant growth promoting endophytes facilitates accumulation of saponin under moderate drought stress

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    摘要:
    目的:滇重樓是中國云南重要的藥用植物資源之一，通常需要6-8年才能采收。因此，迫切需要找到一種既能縮短其生長年限，又能提高其品質(zhì)的方法。在本研究中，我們研究了干旱脅迫下叢枝菌根真菌（AMF）和植物根際促生細(xì)菌(PGPE)聯(lián)合處理對(duì)其皂苷積累的影響。 方法:在干旱脅迫下，用叢枝菌根真菌和植物根際促生細(xì)菌混合侵染滇重樓。測定其總皂苷含量及形態(tài)、生理、生化指標(biāo)。通過同源比較，從轉(zhuǎn)錄組數(shù)據(jù)中獲得與皂苷合成相關(guān)的UGTs基因，進(jìn)行RT-PCR和系統(tǒng)發(fā)育分析。 結(jié)果：不論水分條件如何，AMF處理均可侵染滇重樓根部，但AMF+PGPE處理可降低AMF侵染率。不同水分條件下，單獨(dú)接種AMF與接種AMF+PGPE處理的滇重樓株高、地上鮮重和地下鮮重差異不顯著，但與未接種處理相比，接種處理顯著提高了滇重樓的株高。不同水分條件下，單獨(dú)接種AMF顯著提高了滇重樓的凈光合速率、氣孔導(dǎo)度和蒸騰速率，而雙接種AMF + PGPE顯著提高了胞間CO2濃度和葉綠素?zé)晒鈪?shù)(Fv/Fm)。在不同水分處理下，單獨(dú)接種AMF處理的脯氨酸含量最高，而與正常水分條件相比，在中度干旱條件下，接種AMF + PGPE可顯著提高植株中ABA和IAA的含量。不同水分條件下，接種AMF + PGPE處理提高了滇重樓根莖中N、P、K的比例。與正常水份條件相比，在中度干旱脅迫下，AMF + PGPE處理顯著提高了滇重樓皂苷I、II、VII和總皂苷的含量，同時(shí)法尼基焦磷酸合成酶(FPPS)、香葉基焦磷酸合成酶(GPPS)、環(huán)阿屯醇合成酶(CAS)和角鯊烯環(huán)氧化酶(SE1)基因顯著上調(diào)。皂苷含量與CAS、GPPS和SE1的表達(dá)呈正相關(guān)。皂苷VI含量與糖基轉(zhuǎn)移酶UGT 010922基因表達(dá)顯著相關(guān)，皂苷II含量與UGT010935基因表達(dá)顯著相關(guān)。 結(jié)論：在中度干旱條件下，AMF + PGPE處理更有利于滇重樓激素含量、養(yǎng)分吸收和總皂苷含量的增加，AMF + PGPE處理可在不同程度上調(diào)節(jié)一條或多條甾體皂苷合成通路中關(guān)鍵基因和UGTs基因的表達(dá)，從而刺激滇重樓根莖中甾體皂苷的合成和積累。AMF和PGPE聯(lián)合接種以及適當(dāng)?shù)耐寥栏珊禇l件有利于提高滇重樓品質(zhì)。

    Abstract:
    Objective: Paris polyphylla var. yunnanensis, one of the important medicinal plant resources in Yunnan, China, usually takes 6–8 years to be harvested. Therefore, it is urgent to find a method that can not only shorten its growth years, but also improve its quality. In this study, we examined the effects of a combination treatment of arbuscular mycorrhizal fungi (AMF) and plant growth-promoting endophytes (PGPE) and drought stress on the accumulation of saponins in it. Methods: P. polyphylla var. yunnanensis was infected with a mixture of AMF and PGPE under drought stress. The content of saponins, as well as morphological, physiological, and biochemical indicators, were all measured. The UGTs gene related to saponin synthesis was obtained from transcriptome data by homologous comparison, which were used for RT-PCR and phylogenetic analysis. Results: Regardless of water, AMF treatment could infect the roots of P. polyphylla var. yunnanensis, however double inoculation with AMF and PGPE (AMF + PGPE) would reduce the infection rate of AMF. Plant height, aboveground and underground fresh weight did not differ significantly between the single inoculation AMF and the double inoculation treatment under different water conditions, but the inoculation treatment significantly increased the plant height of P. polyphylla var. yunnanensis compared to the noninoculation treatment. Single inoculation with AMF considerably increased the net photosynthetic rate, stomatal conductance, and transpiration rate of P. polyphylla var. yunnanensis leaves under various water conditions, but double inoculation with AMF + PGPE greatly increased the intercellular CO2 concentration and chlorophyll fluorescence parameter (Fv/Fm). Under diverse water treatments, single inoculation AMF had the highest proline content, whereas double inoculation AMF + PGPE may greatly improve the amount of abscisic acid (ABA) and indoleacetic acid (IAA) compared to normal water under moderate drought. Double inoculation AMF + PGPE treatment improved the proportion of N, P, and K in the rhizome of P. polyphylla var. yunnanensis under various water conditions. Under moderate drought stress, AMF + PGPE significantly enhanced the contents of P. polyphylla var. yunnanensis saponins I, II, VII, and total saponins as compared to normal water circumstances. Farnesyl diphosphate synthase (FPPS), Geranyl pyrophosphate synthase (GPPS), Cycloartenol synthase (CAS), and Squalene epoxidase (SE1) were the genes that were significantly up-regulated at the same time. The amount of saponins was favorably linked with the expression of CAS, GPPS, and SE1. Saponin VI content and glycosyl transferase (UGT) 010922 gene expression were found to be substantially associated, as was saponin II content and UGT010935 gene expression. Conclusion: Under moderate drought, AMF + PGPE was more conducive to the increase of hormone content, nutrient absorption, and total saponin content in P. polyphylla var. yunnanensis, and AMF + PGPE could up regulate the expression of key genes and UGTs genes in one or more steroidal saponin synthesis pathways to varying degrees, thereby stimulating the synthesis and accumulation of steroidal saponins in the rhizome of P. polyphylla var. yunnanensis. The combination of AMF and PGPE inoculation, as well as adequate soil drought, reduced the buildup of saponins in P. polyphylla var. yunnanensis and increased its quality.

    關(guān)鍵詞:
    叢枝菌根真菌;基因表達(dá);重樓;植物促生長內(nèi)生菌;甾體皂苷

    Keywords:


    Project Supported:
    This work was supported by grants from Major Special Projects of the Ministry of Science and Technology (No. 2021yfd1000202; 2021YFD1601003), the Key R & D program of Yunnan Province (No. 202103AC100003) and National Natural Science Foundation of China (No. 31860075).

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