曬黃煙調(diào)制期煙葉真菌多樣性研究
發(fā)布時間:2019-08-25 來源: 人生感悟 點擊:
摘 要:為揭示曬黃煙調(diào)制過程中煙葉真菌的多樣性,采用雙平板純培養(yǎng)分離和ITS序列系統(tǒng)發(fā)育分析的方法,對調(diào)制前期、中期和結(jié)束期煙葉樣品中的真菌進(jìn)行了分離和初步歸類,將代表性菌株進(jìn)行ITS序列測定和系統(tǒng)發(fā)育分析,研究了曬黃煙調(diào)制過程中純培養(yǎng)真菌的群落結(jié)構(gòu)組成和動態(tài)變化。結(jié)果顯示,從曬黃煙調(diào)制期煙葉中共分離獲得126株真菌,下部和上部煙葉分離的菌株數(shù)多于中部煙葉,調(diào)制中期分離的菌株數(shù)少于調(diào)制前期和調(diào)制結(jié)束期;這些菌株分布在2門、5綱、10目、14科和17屬;93.65%的菌株屬于Ascomycota門,Epicoccum、Arthrinium和Fusarium為優(yōu)勢屬,分別占23.02%、13.49%和11.90%。研究表明,曬黃煙在調(diào)制期間,煙葉真菌類群多樣性比較豐富,在不同煙葉部位的分布有所不同,并隨著調(diào)制的進(jìn)行而發(fā)生動態(tài)變化。
關(guān)鍵詞:曬黃煙;調(diào)制;真菌;多樣性
中圖分類號:S435.72 文章編號:1007-5119(2017)06-0012-08 DOI:10.13496/j.issn.1007-5119.2017.06.002
Abstract: To reveal the fungal diversity of yellow sun-cured tobacco during curing, the composition and dynamics of fungal communities were analyzed by a combination of pure cultivation and molecular identification techniques. Cultivable fungi were isolated from the early, middle and late stages of curing. The representative strains were then phylogenetically analyzed based on ITS sequences. The results showed that a total of 126 fungal strains were isolated. The strains isolated from the middle leaves were relatively less compared with those from the bottom and upper leaves. In addition, the tobacco leaves seemed to contain more fungi at the early and late stages comparing to those at the middle stage of curing. The 126 fungal strains were distributed in 2 phyla, 5 classes, 10 orders, 14 families and 17 genera. Ascomycota was the dominant phylum, with 93.65% of the isolates belonging to this phylum, and Epicoccum, Arthrinium and Fusarium were the dominant genera, accounting for 23.02%, 13.49% and 11.90% of the isolates, respectively. It was found that the fungal diversity of yellow sun-cured tobacco leaves during curing was relatively abundant, and the fungal composition varied in different leaf positions and dynamically changed as the curing process proceeded.
Keywords: yellow sun-cured tobacco; curing; fungi; diversity
成熟的煙葉采收后,大量的微生物,包括細(xì)菌、真菌、酵母和放線菌等,仍然存在于煙葉的表面或組織內(nèi)。在煙葉的調(diào)制過程中,這些微生物通過分解煙葉中的糖類和蛋白質(zhì)等物質(zhì)來獲取自身生長所需的營養(yǎng)和能量,并將這些大分子物質(zhì)分解為酚類、醇類、酯類等小分子物質(zhì),微生物的代謝活動
對煙葉化學(xué)成分的組成和煙葉品質(zhì)的形成都會產(chǎn)生重要的影響[1]。煙葉細(xì)菌的多樣性及其對煙葉品質(zhì)形成的影響研究報道較多[2-7]。真菌是煙葉微生物的另一大類群,數(shù)量僅次于細(xì)菌[6]。與細(xì)菌相比,目前對煙草真菌種群和結(jié)構(gòu)的研究比較少,主要是采用純培養(yǎng)方法對一些烤煙煙草內(nèi)生真菌進(jìn)行了分離鑒定[8-11]。煙草真菌具有有利和有害兩個方面的作用,如防治煙草病蟲害[8]、降低煙堿含量[12-14]、改善煙葉品質(zhì)[15-17]、引起煙草病害[18]和煙葉腐爛變質(zhì)等[19-21]。
曬黃煙是我國特有的一種煙葉類型,幾百年的栽培和種植使之形成了獨特的煙葉品質(zhì),是我國發(fā)展具有獨特香氣風(fēng)格和口味特征的中式卷煙的優(yōu)質(zhì)原料。本文采用純培養(yǎng)分離和ITS序列系統(tǒng)發(fā)育分析方法,研究曬黃煙煙葉晾曬調(diào)制期真菌群落結(jié)構(gòu)的組成和動態(tài)變化,為人工調(diào)控并改進(jìn)曬黃煙煙葉調(diào)制方式,提升煙葉品質(zhì)提供理論指導(dǎo)。
1 材料與方法
1.1 試驗材料
供試樣品取自曬黃煙云曬1號調(diào)制過程中的煙葉,該品種種植于云南盈江弄璋曬黃煙基地。
1.2 方法
1.2.1 煙葉調(diào)制和取樣 云曬1號煙葉的調(diào)制和取樣方法參照倪紅梅等[7]的方法,2013年3月至4月期間,依次收集下、中和上3個部位的新鮮煙葉(調(diào)制第1天)、調(diào)制中期煙葉(調(diào)制第7天)和調(diào)制結(jié)束煙葉(調(diào)制第13天)。
相關(guān)熱詞搜索:真菌 煙葉 多樣性 調(diào)制 研究
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