杏鮑菇液態菌種應用於出菇之研究

Applied of liquid spawn on fruiting of Pleurotus eryngii

陳彥廷(Yan-Ting Chen)

指導教授 ： 詹効松 ； 石信德

朝陽科技大學/理工學院/應用化學系生化科技碩博士班/碩士(2016年)

摘要

本研究擬建立杏鮑菇液態菌種之活性分析指標，以杏鮑菇菌絲體於CS基礎培養基與本實驗室所研發之最適化液態培養基進行液態菌種培養，分析其纖維素與木質素分解酵素(cellulase, laccase)活性、液態菌種接入木屑太空包之走菌速率、出菇率及子實體產量為評斷依據，探討金屬離子添加對於杏鮑菇液態菌種酵素活性及杏鮑菇子實體產量之影響。結果顯示於CS基礎培養基添加1 mg/L硫酸銅有助於菌絲體生質量之增加，於培養第10天可獲得最高生質量3.00±0.49 g/L，高於對照組2.27±0.43 g/L；硫酸銅添加濃度範圍於1至10 mg/L時有助於提升菌絲體生質量、纖維素和木質素分解酵素活性；本研究為了可快速得到液態菌種應用於菇蕈類子實體生產實務上之參數，將木屑太空包(1 kg)與依等比例縮小（1/50, w/w) 配製之木屑試管進行杏鮑菇液態菌種接種、菌絲走菌速率、添加物試驗（出菇前）以及出菇等試驗，並與傳統之固態菌種進行比較，俾利於收集相關參數並應用於大規模之生產實務。結果發現以杏鮑菇液態菌種於不同稀釋（1至40倍 ）處理及添加銅、錳、鋅等離子後分別接種於木屑試管和太空包，觀察其菌絲體分別於20和28天走菌完成，其值均優於固態菌種之試驗組(25和35天)。在出菇前添加添營養液至木屑試管和太空包內，由出菇率及子實體產量變化可發現此方式對於子實體產量提升具有顯著性。以稀釋40倍之液態菌種（最適培養基）接種至木屑太空包，於出菇前添加20 ml 營養液之試驗組，子實體產量(296.03±6.72 g/bag）顯著高於控制組(230.64±20.55 g/bag)。綜合上述，本研究以纖維素及木質素分解酵素活性之分析與杏鮑菇子實體產量，可間接證明液態菌種應用於木屑太空包生產杏鮑菇優於固態菌種，此外，杏鮑菇液態菌種添加銅、錳離子可縮短木屑太空包菌絲走菌完成時間並提升杏鮑菇子實體產量。期許本研究之數據有助於杏鮑菇子實體生產實務與產業發展。

關鍵字

杏鮑菇 ； 液態菌種培養 ； 纖維素分解酵素 ； 木質素分解酵素 ； 木屑試管 ； 木屑太空包 ； 子實體

並列摘要

This research intends to establish an active analysis target of liquid spawn for Pleurotus eryngii. P. eryngii was cultured in czapek solution based medium and the optimum cultivation liquid medium. Then, we analyzed the enzyme activity in the cultured mycelia. Considering the rate of mycelium growth and the yield of fruitingbodies, we aimed to improve the enzyme activity in P. eryngii strains. With a single factor of metal ions additive test, enhanced enzyme activity was observed in strain cultured in liquid medium. The results showed that a concentration of 1 mg/L copper(II) sulfate enhanced the biomass. The highest biomass (3.00 ± 0.49 g/L) was recorded at 10th day of culture. The biomass was higher compared to the control group (2.27 ± 0.43 g/L). A concentration of 1-10 mg/L copper(II) sulfate promoted the enzyme activity (cellulase and Laccase). To shorten the cultured time, we adjusted the spawn growth and fruiting conditions to the sawdust tube in place of bag cultivation. The results showed that the cultivation in liquid and dilution folds of copper, manganese and zinc ions in the sawdust tubes were better than the bag. The rate of mycelium growth in liquid culture (20 and 28 days) were better than the solid culture (25 and 35 days). It was found that addition of agents (water and organic nitrogen sources) into the sawdust tubes and space packages could enhance the yield of fruitingbodies significantly. The yield of fruitingbodies was 317.90±7.47g/bag by adding the diluting 40-folds optimization medium liquid strains in the 20mL water testing group. This increased to 341.16±2.86g/bag by adding the diluting 10-folds optimization medium liquid strains in the 20mL nitrogen sources testing group and was the highest biomass.
In conclusion, we could prove that the strains in liquid culture were better than the strain in solid medium considering the space packages by the analysis of cellulolytic and ligninolytic enzyme activities. Also, addition of Copper and Manganese ions shortened the rate of mycelium growth and improved the yield of P. eryngii fruitingbodies. The results of our study hence could be helpful in the yield of fruiting bodies.

並列關鍵字

Pleurotus eryngii ； liquid spawn culture ； cellulolytic enzymes ； ligninolytic enzyme ； sawdust tube ； space package ； fruiting body

參考文獻

石信德，｢食藥用菇類液體菌種栽培技術之潛力｣，農業生技產業季刊，NO.23，第16-21頁 (2010)。

李澤宏、王均琍，｢六種菇菌纖維素分解酵素與木聚醣分解酵素活性之分析｣，高雄區農業改良場研究彙報，第20卷，第1期，第26-33頁(2011)。

林彥良、王升陽、曲芳華，｢紅檜 Caffeoyl-CoA 3-O-methyltransferase基因之選殖及其轉基因菸草木質素分析｣，中華林學季刊，第41卷，第3期，第323-338頁 (2008)。

陳永龍、陳載永、吳志鴻，｢淺談真菌降解木材機制與環境友善型木材防腐劑｣，中華林學季刊，第43卷，第一期，第181-189頁 (2010)。

陳美杏、李瑋崧、吳寬澤、簡宣裕、呂昀陞，｢杏鮑菇廢棄基質在菇類栽培之應用｣，台灣農業研究，第62期，第126-136頁 (2013)。