氮源对蛹虫草生长及类胡萝卜素产生的影响
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摘要
【背景】蛹虫草是一种珍稀食药用菌,类胡萝卜素不仅为其重要活性成分,而且影响子实体的外观品相,但是类胡萝卜素产生的影响因素不明。【目的】揭示氮源对蛹虫草生长和类胡萝卜素产生的影响。【方法】测定不同氮源培养基中菌株生长速度、分生孢子产生及类胡萝卜含量,筛选菌株生长的最适氮源,进一步研究不同浓度的氮源对蛹虫草生长及类胡萝卜素产生的影响,并测定不同光照条件下氮源浓度对蛹虫草子实体类胡萝卜素产生的影响。【结果】蛹虫草在不同的氮源培养基中菌落形态和类胡萝卜素产生存在明显差异。麦麸和黄豆粉培养基中菌株生长速度最快,但是菌落稀疏,正面分别呈现荧光黄色和极微弱红色;蛋白胨和酵母提取物培养基中菌落致密,产孢量极显著高于其它氮源培养基(P<0.01),菌落正面为橙黄色;甘氨酸和柠檬酸为氮源时完全没有色素产生,其它无机和氨基酸氮源培养基平板背面有微量色素产生。固体和液体静置培养条件下均发现蛋白胨浓度在0-3%的范围内,随着浓度的增加,类胡萝卜素含量增加;子实体栽培中不同的氮源浓度均表现为蓝光光照条件下类胡萝卜素含量显著高于白光;而在白光和蓝光光照条件下均表现为蛋白胨浓度为1%时类胡萝卜素含量最高,分别为2 809.38±386.24μg/g和4 093.75±518.37μg/g。【结论】氮源种类和浓度显著影响蛹虫草类胡萝卜素的产生,蓝光光照和1%蛋白胨浓度为子实体类胡萝卜素产生的最佳条件,这为栽培富含类胡萝卜素的蛹虫草子实体提供了试验依据。
[Background] Cordyceps militaris is one of the famous medicinal fungi. Carotenoid is not only an important active component but also affects the appearance of fruiting body. However, the factors regulating its production are unknown. [Objective] To reveal the effects of nitrogen sources on the growth and carotenoid production of C. militaris. [Methods] The growth rate, conidia and carotenoid production on media with different nitrogen sources were determined and the optimum nitrogen sources were selected. The effects of nitrogen concentration on carotenoid production of C. militaris fruiting body under different light conditions were further determined. [Results] There were significant differences in growth and carotenoid production of C. militaris on media with different nitrogen sources. The growth rate was the fastest in wheat bran and soybean powder media, showing as fluorescent yellow and very weak red, respectively. The conidia produced in peptone and yeast extract media were significantly more than that in other nitrogen sources(P<0.01), and the colony was orange. There was no pigment production in inorganic nitrogen source media except for trace pigments on the back of plate of glutamic acid and ammonium chloride. It was found that the carotenoid content increased with the increase of peptone concentration in the range of 0-3%. Blue light was more suitable for the carotenoid production in fruiting body than white light. The carotenoid contents were the highest with the peptone concentration of 1% under both white and blue light conditions, 2 809.38±386.24 μg/g and 4 093.75±518.37 μg/g respectively. [Conclusion] Nitrogen source and its concentration significantly affected the carotenoid production in C. militaris. Blue light and peptone concentration of 1% were the best conditions for carotenoid production in fruiting bodies. This study will be helpful for cultivating C. militaris fruiting body rich in carotenoid.
[Background] Cordyceps militaris is one of the famous medicinal fungi. Carotenoid is not only an important active component but also affects the appearance of fruiting body. However, the factors regulating its production are unknown. [Objective] To reveal the effects of nitrogen sources on the growth and carotenoid production of C. militaris. [Methods] The growth rate, conidia and carotenoid production on media with different nitrogen sources were determined and the optimum nitrogen sources were selected. The effects of nitrogen concentration on carotenoid production of C. militaris fruiting body under different light conditions were further determined. [Results] There were significant differences in growth and carotenoid production of C. militaris on media with different nitrogen sources. The growth rate was the fastest in wheat bran and soybean powder media, showing as fluorescent yellow and very weak red, respectively. The conidia produced in peptone and yeast extract media were significantly more than that in other nitrogen sources(P<0.01), and the colony was orange. There was no pigment production in inorganic nitrogen source media except for trace pigments on the back of plate of glutamic acid and ammonium chloride. It was found that the carotenoid content increased with the increase of peptone concentration in the range of 0-3%. Blue light was more suitable for the carotenoid production in fruiting body than white light. The carotenoid contents were the highest with the peptone concentration of 1% under both white and blue light conditions, 2 809.38±386.24 μg/g and 4 093.75±518.37 μg/g respectively. [Conclusion] Nitrogen source and its concentration significantly affected the carotenoid production in C. militaris. Blue light and peptone concentration of 1% were the best conditions for carotenoid production in fruiting bodies. This study will be helpful for cultivating C. militaris fruiting body rich in carotenoid.
引文
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[7]Yan XT,Bao HY,Bau T.Isolation and identification of one natural pigment from cultured Cordyceps militaris[J].Mycosystema,2010,29(5):777-781(in Chinese)闫喜涛,包海鹰,图力古尔.人工培养蛹虫草中一种天然色素的分离和结构鉴定[J].菌物学报,2010,29(5):777-781
[8]Chen C.Chemical study of cultured Cordyceps militaris[D].Changchun:Master’s Thesis of Jilin Agricultural University,2012(in Chinese)陈策.人工蛹虫草的化学成分研究[D].长春:吉林农业大学硕士学位论文,2012
[9]Dong JZ,Wang SH,Ai XR,et al.Composition and characterization of cordyxanthins from Cordyceps militaris fruit bodies[J].Journal of Functional Foods,2013,5(3):1450-1455
[10]Zhang XJ,Chen XX,Tang J,et al.Study on the stability of Cordyceps militaris yellow pigment A1[J].Science and Technology of Food Industry,2012,33(18):128-130,134(in Chinese)张晓君,陈雪香,唐健,等.蛹虫草黄色素A1稳定性研究[J].食品工业科技,2012,33(18):128-130,134
[11]Yin L,Lin JF,Qian J,et al.Optimization of the liquid culture medium composition for high carotenoid production by Cordyceps militaris and the investigation on the improvement of carotenoid extraction[J].Science and Technology of Food Industry,2017,38(18):79-84(in Chinese)殷林,林俊芳,钱璟,等.高产类胡萝卜素的蛹虫草液体培养基优化及其提叐工艺研究[J].食品工业科技,2017,38(18):79-84
[12]Yang T,Dong CH.Photo morphogenesis and photo response of the blue-light receptor gene Cmwc-1 in different strains of Cordyceps militaris[J].FEMS Microbiology Letters,2014,352(2):190-197
[13]Shen JL,Jin HY,Gao Y,et al.Two-step fermentation of carotenoid by blue light induction in Cordyceps militaris L.[J].China Biotechnology,2013,33(2):83-87(in Chinese)沈俊良,金华燕,高雅,等.蛹虫草蓝光诱导两步法収酵产类胡萝卜素[J].中国生物工程杂志,2013,33(2):83-87
[14]Wu SM,Zhou LH,Shi W,et al.Influence of light intensity on the growth and development of carotene content and body of Cordyceps militaris[J].Guangdong Agricultural Sciences,2013,40(7):24-25,45(in Chinese)吴铄溟,周礼红,史炜,等.光照对蛹虫草菌丝体生长収育及类胡萝卜素含量的影响[J].广东农业科学,2013,40(7):24-25,45
[15]Lian TT,Dong CH,Yang T,et al.Effects of blue light on the growth and bioactive compound production of Cordyceps militaris[J].Mycosystema,2014,33(4):838-846(in Chinese)廉添添,董彩虹,杨涛,等.蓝光光照对蛹虫草子实体生长和主要活性成分的影响[J].菌物学报,2014,33(4):838-846
[16]Zhang JJ,Wang F,Liu KB,et al.Heat and light stresses affect metabolite production in the fruit body of the medicinal mushroom Cordyceps militaris[J].Applied Microbiology and Biotechnology,2018,102(10):4523-4533
[17]Tudzynski B.Nitrogen regulation of fungal secondary metabolism in fungi[J].Frontiers in Microbiology,2014,5:656
[18]Li F,Wang Z,Bian WY,et al.Effects of five kinds nitrogen sources on the growth and quality of Cordyceps militaris[J].Fujian Agricultural Science and Technology,2018(1):4-6(in Chinese)李菲,王忠,卞文印,等.5种氮源对蛹虫草生长及其质量的影响[J].福建农业科技,2018(1):4-6
[19]Fang HZ,Xiao XM.Effects of different nitrogen sources on the main active components of Cordyceps militaris[J].Journal of Jingchu University of Technology,2016,31(6):5-10(in Chinese)方华舟,肖习明.不同氮源对蛹虫草主要活性成分的影响及觃律研究[J].荆楚理工学院学报,2016,31(6):5-10
[20]Jian LR,Du ST.Effect of nitrogen and inorganic salt on production of stroma,cordycepin and adenosine in Cordyceps militaris[J].Northern Horticulture,2016(19):163-167(in Chinese)简利茹,杜双田.氮源和无机盐对蛹虫草子座产量及虫草素和腺苷的影响[J].北方园艺,2016(19):163-167
[21]Gu YY,Zhang YP,Chen CJ,et al.Effects of carbon and nitrogen sources in culture medium on growth amount of Cordyceps militaris hyphae[J].Shandong Agricultural Sciences,2008(7):77-79(in Chinese)顾寅钰,张亚平,陈传杰,等.蛹虫草培养基碳源和氮源对菌丝生长量的影响[J].山东农业科学,2008(7):77-79
[22]Zhou B,Pu YW,Zhu MJ,et al.Effects of nitrogen sources on Monascus yellow pigment production by Monascus mutant[J].Modern Food Science and Technology,2008,24(2):123-127(in Chinese)周波,浦跃武,朱明军,等.氮源对红曲霉突叒株产黄色素的影响[J].现代食品科技,2008,24(2):123-127
[23]Yang CL,Yang XJ,He ZG,et al.Effects of carbon and nitrogen sources on Monascus purpureus S in its secreting pigment[J].Fujian Journal of Agricultural Sciences,2011,26(5):832-836(in Chinese)杨成龙,杨晓君,何志刚,等.碳氮源对红曲霉S产色素的影响[J].福建农业学报,2011,26(5):832-836
[24]Zhu ZY,Han D,Pang W.Study on the producting conditions of pigment by Paecilomyces sinensis[J].Food Research and Development,2017,38(2):190-195(in Chinese)朱振元,韩丹,庞伟.中国拟青霉产色素収酵条件的研究[J].食品研究与开収,2017,38(2):190-195
[25]Guo MM,Guo SP,Yang HJ,et al.Comparison of major bioactive compounds of the caterpillar medicinal mushroom,Cordyceps militaris(Ascomycetes),fruiting bodies cultured on wheat substrate and pupae[J].International Journal of Medicinal Mushrooms,2016,18(4):327-336
[26]Rodriguez-Amaya DB.Quantitative analysis,in vitro assessment of bioavailability and antioxidant activity of food carotenoids--a review[J].Journal of Food Composition and Analysis,2010,23(7):726-740
[27]Zhao N,Yuan XL,Chen J,et al.Effect of different carbon and nitrogen sources on mycelia growth of Antrodia cinnamomea[J].Journal of West China Forestry Science,2016,45(4):7-12(in Chinese)赵能,原晓龙,陈剑,等.不同碳氮源对牛樟芝菌丝体生长的影响[J].西部林业科学,2016,45(4):7-12
[28]Gmoser R,Ferreira JA,Lennartsson PR,et al.Filamentous ascomycetes fungi as a source of natural pigments[J].Fungal Biology and Biotechnology,2017,4:4
[2]Xia YL,Luo FF,Shang YF,et al.Fungal cordycepin biosynthesis is coupled with the production of the safeguard molecule pentostatin[J].Cell Chemical Biology,2017,24(12):1479-1489
[3]Fu MJ.Study on the carotenoid produced from Cordyceps militaris L.[J].Journal of Food Science and Biotechnology,2005,24(5):107-110(in Chinese)付鸣佳.蛹虫草产类胡萝卜素的研究[J].食品与生物技术学报,2005,24(5):107-110
[4]Yang T,Sun JD,Lian TT,et al.Process optimization for extraction of carotenoids from medicinal caterpillar fungus,Cordyceps militaris(Ascomycetes)[J].International Journal of Medicinal Mushrooms,2014,16(2):125-135
[5]Fu MJ,Wang XJ,Huang WF.Blue light induced carotenoid accumulation in Cordyceps militaris L.[J].Microbiology China,2005,32(5):24-28(in Chinese)付鸣佳,王小菁,黄文芳.蓝光诱导蛹虫草菌丝类胡萝卜素的积累[J].微生物学通报,2005,32(5):24-28
[6]Zhang ZJ,Jiang XL,Mou HJ,et al.Study of extraction conditions on the carotenoid produced from Cordyceps militaris[J].Food Science and Technology,2007(4):99-103(in Chinese)张志军,江晓路,牟海津,等.蛹虫草类胡萝卜素提叐工艺的研究[J].食品科技,2007(4):99-103
[7]Yan XT,Bao HY,Bau T.Isolation and identification of one natural pigment from cultured Cordyceps militaris[J].Mycosystema,2010,29(5):777-781(in Chinese)闫喜涛,包海鹰,图力古尔.人工培养蛹虫草中一种天然色素的分离和结构鉴定[J].菌物学报,2010,29(5):777-781
[8]Chen C.Chemical study of cultured Cordyceps militaris[D].Changchun:Master’s Thesis of Jilin Agricultural University,2012(in Chinese)陈策.人工蛹虫草的化学成分研究[D].长春:吉林农业大学硕士学位论文,2012
[9]Dong JZ,Wang SH,Ai XR,et al.Composition and characterization of cordyxanthins from Cordyceps militaris fruit bodies[J].Journal of Functional Foods,2013,5(3):1450-1455
[10]Zhang XJ,Chen XX,Tang J,et al.Study on the stability of Cordyceps militaris yellow pigment A1[J].Science and Technology of Food Industry,2012,33(18):128-130,134(in Chinese)张晓君,陈雪香,唐健,等.蛹虫草黄色素A1稳定性研究[J].食品工业科技,2012,33(18):128-130,134
[11]Yin L,Lin JF,Qian J,et al.Optimization of the liquid culture medium composition for high carotenoid production by Cordyceps militaris and the investigation on the improvement of carotenoid extraction[J].Science and Technology of Food Industry,2017,38(18):79-84(in Chinese)殷林,林俊芳,钱璟,等.高产类胡萝卜素的蛹虫草液体培养基优化及其提叐工艺研究[J].食品工业科技,2017,38(18):79-84
[12]Yang T,Dong CH.Photo morphogenesis and photo response of the blue-light receptor gene Cmwc-1 in different strains of Cordyceps militaris[J].FEMS Microbiology Letters,2014,352(2):190-197
[13]Shen JL,Jin HY,Gao Y,et al.Two-step fermentation of carotenoid by blue light induction in Cordyceps militaris L.[J].China Biotechnology,2013,33(2):83-87(in Chinese)沈俊良,金华燕,高雅,等.蛹虫草蓝光诱导两步法収酵产类胡萝卜素[J].中国生物工程杂志,2013,33(2):83-87
[14]Wu SM,Zhou LH,Shi W,et al.Influence of light intensity on the growth and development of carotene content and body of Cordyceps militaris[J].Guangdong Agricultural Sciences,2013,40(7):24-25,45(in Chinese)吴铄溟,周礼红,史炜,等.光照对蛹虫草菌丝体生长収育及类胡萝卜素含量的影响[J].广东农业科学,2013,40(7):24-25,45
[15]Lian TT,Dong CH,Yang T,et al.Effects of blue light on the growth and bioactive compound production of Cordyceps militaris[J].Mycosystema,2014,33(4):838-846(in Chinese)廉添添,董彩虹,杨涛,等.蓝光光照对蛹虫草子实体生长和主要活性成分的影响[J].菌物学报,2014,33(4):838-846
[16]Zhang JJ,Wang F,Liu KB,et al.Heat and light stresses affect metabolite production in the fruit body of the medicinal mushroom Cordyceps militaris[J].Applied Microbiology and Biotechnology,2018,102(10):4523-4533
[17]Tudzynski B.Nitrogen regulation of fungal secondary metabolism in fungi[J].Frontiers in Microbiology,2014,5:656
[18]Li F,Wang Z,Bian WY,et al.Effects of five kinds nitrogen sources on the growth and quality of Cordyceps militaris[J].Fujian Agricultural Science and Technology,2018(1):4-6(in Chinese)李菲,王忠,卞文印,等.5种氮源对蛹虫草生长及其质量的影响[J].福建农业科技,2018(1):4-6
[19]Fang HZ,Xiao XM.Effects of different nitrogen sources on the main active components of Cordyceps militaris[J].Journal of Jingchu University of Technology,2016,31(6):5-10(in Chinese)方华舟,肖习明.不同氮源对蛹虫草主要活性成分的影响及觃律研究[J].荆楚理工学院学报,2016,31(6):5-10
[20]Jian LR,Du ST.Effect of nitrogen and inorganic salt on production of stroma,cordycepin and adenosine in Cordyceps militaris[J].Northern Horticulture,2016(19):163-167(in Chinese)简利茹,杜双田.氮源和无机盐对蛹虫草子座产量及虫草素和腺苷的影响[J].北方园艺,2016(19):163-167
[21]Gu YY,Zhang YP,Chen CJ,et al.Effects of carbon and nitrogen sources in culture medium on growth amount of Cordyceps militaris hyphae[J].Shandong Agricultural Sciences,2008(7):77-79(in Chinese)顾寅钰,张亚平,陈传杰,等.蛹虫草培养基碳源和氮源对菌丝生长量的影响[J].山东农业科学,2008(7):77-79
[22]Zhou B,Pu YW,Zhu MJ,et al.Effects of nitrogen sources on Monascus yellow pigment production by Monascus mutant[J].Modern Food Science and Technology,2008,24(2):123-127(in Chinese)周波,浦跃武,朱明军,等.氮源对红曲霉突叒株产黄色素的影响[J].现代食品科技,2008,24(2):123-127
[23]Yang CL,Yang XJ,He ZG,et al.Effects of carbon and nitrogen sources on Monascus purpureus S in its secreting pigment[J].Fujian Journal of Agricultural Sciences,2011,26(5):832-836(in Chinese)杨成龙,杨晓君,何志刚,等.碳氮源对红曲霉S产色素的影响[J].福建农业学报,2011,26(5):832-836
[24]Zhu ZY,Han D,Pang W.Study on the producting conditions of pigment by Paecilomyces sinensis[J].Food Research and Development,2017,38(2):190-195(in Chinese)朱振元,韩丹,庞伟.中国拟青霉产色素収酵条件的研究[J].食品研究与开収,2017,38(2):190-195
[25]Guo MM,Guo SP,Yang HJ,et al.Comparison of major bioactive compounds of the caterpillar medicinal mushroom,Cordyceps militaris(Ascomycetes),fruiting bodies cultured on wheat substrate and pupae[J].International Journal of Medicinal Mushrooms,2016,18(4):327-336
[26]Rodriguez-Amaya DB.Quantitative analysis,in vitro assessment of bioavailability and antioxidant activity of food carotenoids--a review[J].Journal of Food Composition and Analysis,2010,23(7):726-740
[27]Zhao N,Yuan XL,Chen J,et al.Effect of different carbon and nitrogen sources on mycelia growth of Antrodia cinnamomea[J].Journal of West China Forestry Science,2016,45(4):7-12(in Chinese)赵能,原晓龙,陈剑,等.不同碳氮源对牛樟芝菌丝体生长的影响[J].西部林业科学,2016,45(4):7-12
[28]Gmoser R,Ferreira JA,Lennartsson PR,et al.Filamentous ascomycetes fungi as a source of natural pigments[J].Fungal Biology and Biotechnology,2017,4:4