榆耳新品种选育及营养生理特性研究
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摘要
榆耳(Gloeostereum incamatum S.Ito et Imai),食之似海参,营养丰富,具有医治肠胃疾病的药用价值,是著名的食药用菌。近些年已经由我国率先人工栽培成功,成为东北的主要食用菌之一。但是,在生产中多直接以野生菌株为菌种、凭经验进行栽培管理,品质与产量很不稳定。为保持我国在榆耳栽培方面的领先地位,关于榆耳新品种选育及栽培生理方面的研究工作已迫在眉睫。因此,本试验对通过系选法选育出来的、产量品质均表现出超亲遗传的C-1、C-2、C-3等3个优良菌株进行了进一步筛选,选育玉米芯培养料专用型榆耳新品种,优化最佳培养料配方,并对榆耳胞外酶活性、营养利用规律等营养生理特性进行系统研究,旨在为我国东北等玉米产区的榆耳生产提供玉米芯培养料专用型新品种及配套栽培技术,为榆耳标准化、高效化栽培奠定生理学基础。具体试验结果如下:
(1)通过对峙培养,3个供试菌株之间均产生了明显的拮抗线,说明供试菌株为生理特性各不相同的独立菌株。
(2)对3个供试菌株进行菌丝培养特性的研究表明,各供试菌株的适宜生长温度为10℃至35℃之间,最适宜生长温度为25℃;综合生长速度、满皿时间及菌丝长势,以C-1较佳。
(3)3个供试菌株中C-1对玉米芯培养料的适应性最强。发菌期间,C-1的染菌率较低,仅为3%,比其它供试菌株低3%～5%;菌丝茁壮浓密,蔓延速度快,50d满袋,提早3～7d。子实体发育阶段,C-1显蕾时间明显提前,比其它供试菌株早7～8d;生物学转化率达到66.5%,比其它供试菌株高约15%;子实体质量明显优于其它供试菌株,个大、肉厚、颜色鲜艳(粉红色)、菇形似花。
(4)通过对榆耳胞外酶活性研究表明,榆耳具有完整的胞外酶系,能够降解培养料中的木质纤维素复合体中的全部组分;但不同培养料中的榆耳菌丝分泌的胞外酶活性在其生长发育的各阶段表现出一定差异,但酶活性变化的总体趋势基本相同。纤维素酶系之间具有明显的互补的关系,半纤维素酶在子实体形成的生殖生长阶段酶活性较强。木质素酶系在菌丝生长阶段的酶活性较强,随后逐渐下降。
(5)通过对榆耳营养利用规律的研究表明,各个生长阶段榆耳对培养基中的各种营养成分的降解率各不相同。从淀粉酶活性的变化可以看出,淀粉类非木质纤维素物质是菌丝生长前期的主要营养源,木质素的利用主要是在菌丝生长阶段;对半纤维素的利用主要是在生殖生长阶段;而对纤维素的利用贯穿于整个生长阶段。最终纤维素的减少量平均为51.39%,半纤维素的减少量平均为32.22%,木质素的减少量平均为29.16%,可见纤维素是榆耳生长发育的主要营养源,其次是半纤维素,对木质素的利用相对较少。
(6)通过榆耳在不同培养基中的菌丝生长、产量及生物转化率的研究表明,配方D为本试验最适培养基配方,玉米芯60%、硬杂木屑18%、麸皮20%、石膏1%、糖1%。发菌期间,在配方D中的染菌率低,仅为3%,比其他配方低1%～4%;菌丝生长茁壮、浓密,45d满袋,比其他配方提早3～20d;平均产量0.245Kg,生物转化率为81.78%,比其他配方高8～34%。
(7)采用常规营养分析法对不同培养料所获的子实体营养成分进行测定,结果表明:不同配方所获的榆耳子实体水分含量在90%左右,差异不明显;但可溶性蛋白和可溶性糖含量差异较大,配方D中子实体可溶性蛋白9.06mg/100g,比其他配方高约1～2 mg/100g,达到极显著水平;可溶性糖含量为11.96mg/g,比其他配方高约1.5～4 mg/g,也达到极显著水平。不同培养料上收获的子实体颜色也有所差异,添加玉米芯的培养基配方的子实体颜色较浅。
综上所述,本试验筛选出1个玉米芯培养料专用型榆耳新品种,提出了菌丝最适培养温度和最适培养基配方;明确了榆耳具有完整的胞外酶系,纤维素为其主要营养源,半纤维素次之,木质素较少等营养生理特性。
Gloeostereum incamatum S.Ito et Imai is well-known as edible and medical fungi,eaten like sea cucumber,which has rich nourishment and the medicinal value for treatment of stomach disease.Recently, Gloeostereum incamatum has been artificially cultured initially and successfully in China,it has become one of the most important edible fungi in Northeast.But,quality and quantity is not stable due to the directly using wild species and lacking cultivation management experiment.Research on breeding of new variety and cultivation physiology of Gloeostereum incamatum should be developed immediately so as to keep the leading position of culturing management in China.In this paper,three excellent strains of C-1,C-2 and C-3 which output and quality were superior to their parents were selected from many strains and new species of Gloeostereum incamatum which cultivated with corncob culture materials was selected,Optimization of the compost formula,aimed to establish the basis of breeding and cultivation physiology,offer excellent new variety and corresponding cultivation technology in the corn productive region(northeast of China).The experimental results are as follows:
(1) According to antagonism identification,the tested strains were three different physiological characteristic ones which produced possession of specific antagonism line.
(2) The result of mycelium cultural character study showed:the temperature that three tested strains growing was between 10℃～35℃,the most suitable temperature was 25℃.Combining with growth speed, growth time and mycelium growth status showed that C-1 was the best strain.
(3) The result showed:the optimal adaptability to corncob medium was the strain of C-1.In mycelium growing stage,the contamination of C-1 was only 3%that was lower than the other strains 3%～5%.The mycelium was healthier and stronger,it grew faster and the cultivation bag was full of it in 50d which was earlier than the others for 3-7d.In fruit bodies budding period,the appearing bud time of C-1 was shorter than the other strains for 7～8d.Biological conversion ratio of fruit bodies can reach 66.5%which was 15%higher than the others.The fruit bodies were obviously excelled other strains in big size,thick,colorful(pink) and shape-like flower.
(4) The study on extracellular enzyme activity showed:it had complete extracellular enzyme systems, which could make lignocelluloses degradation completely in medium.Extracellular enzyme activity excreted in different medium formula behaved a certain disparity in each growing stage of Gloeostereum incamatum, but the general tendency of enzyme activity was essential same.The cellulose enzyme system had clearly complementary relationship and the activities of hemicellulase were stronger at the reproductive phase.The activity of lignocellulose enzyme system was higher at the mycelial growth stage and then decreased.
(5) The nutrition utilization regularity study showed:degradation ratios of all kinds of nutrition in each growth phase were different,the mostly utilization of nonlignocellulose and lignocellulose were mainly in the stage of mycelium growth;Form the change of amylase activity,starch materials were the main nutrition at the earlier stage,the main utilization of lignin was at the stage of mycelial growth;Hemicellulose was utilized mainly in the reproductive growth phase;But the utilization of cellulose was prolonged the whole growth phase.Finally,the average decrement of cellose was 51.39%,the hemicellulose was 32.22%and the lignin was 29.16%.Therefore,the test showed that cellulose was the major nutrition source of Gloeostereum incamatum,secondly was hemicellulose,lignin was fewer relatively.
(6) Through the study on mycelium growth condition,yield and biological conversion ratio in different medium,we could find that formula D was the most optimal medium which was composed of 60%corncob, 18%saw dust,20%wheat bran,1%calcium sulfate and 1%sucrose.During the period of culturing mycelium, the contamination ratio of formula D was only 3%which was lower than the other formulas by 1%～4%; mycelium was healthier and stronger and growing faster,filling bag in 45d which earlier 3～20d than the others;average yield was 0.245kg,biological conversion was 81.78%which was 8～34%higher than the other formulas.
(7) The nutrition analysis of fruit bodies which were obtained from different medium formulas showed: moisture content of fruit bodies were about 90%,the difference was not significance.But the content of soluble sugar and protein was in significant difference,the soluble protein of formula D was 9.06mg/100g which higher than the others for about 1～2mg/100g,getting a very significant level.The soluble sugar content was 11.96mg/g,which was higher than the others for about 1.5～4mg/g,reached extremely significant level. The color of fruit bodies which obtained from different medium formulas were different,the color of fruit body which obtained from corncob medium was tint.
Consequently,this experiment screened a best strain which was suitable to corncob medium cultivation and advanced the optimal temperature and formula of mycelium.Studying the change pattern of extracellular enzyme activities and cellulose was the major nutrition source of Gloeostereum incamatum,secondly was hemicellulose,lignin was fewer relatively.
(1)通过对峙培养,3个供试菌株之间均产生了明显的拮抗线,说明供试菌株为生理特性各不相同的独立菌株。
(2)对3个供试菌株进行菌丝培养特性的研究表明,各供试菌株的适宜生长温度为10℃至35℃之间,最适宜生长温度为25℃;综合生长速度、满皿时间及菌丝长势,以C-1较佳。
(3)3个供试菌株中C-1对玉米芯培养料的适应性最强。发菌期间,C-1的染菌率较低,仅为3%,比其它供试菌株低3%～5%;菌丝茁壮浓密,蔓延速度快,50d满袋,提早3～7d。子实体发育阶段,C-1显蕾时间明显提前,比其它供试菌株早7～8d;生物学转化率达到66.5%,比其它供试菌株高约15%;子实体质量明显优于其它供试菌株,个大、肉厚、颜色鲜艳(粉红色)、菇形似花。
(4)通过对榆耳胞外酶活性研究表明,榆耳具有完整的胞外酶系,能够降解培养料中的木质纤维素复合体中的全部组分;但不同培养料中的榆耳菌丝分泌的胞外酶活性在其生长发育的各阶段表现出一定差异,但酶活性变化的总体趋势基本相同。纤维素酶系之间具有明显的互补的关系,半纤维素酶在子实体形成的生殖生长阶段酶活性较强。木质素酶系在菌丝生长阶段的酶活性较强,随后逐渐下降。
(5)通过对榆耳营养利用规律的研究表明,各个生长阶段榆耳对培养基中的各种营养成分的降解率各不相同。从淀粉酶活性的变化可以看出,淀粉类非木质纤维素物质是菌丝生长前期的主要营养源,木质素的利用主要是在菌丝生长阶段;对半纤维素的利用主要是在生殖生长阶段;而对纤维素的利用贯穿于整个生长阶段。最终纤维素的减少量平均为51.39%,半纤维素的减少量平均为32.22%,木质素的减少量平均为29.16%,可见纤维素是榆耳生长发育的主要营养源,其次是半纤维素,对木质素的利用相对较少。
(6)通过榆耳在不同培养基中的菌丝生长、产量及生物转化率的研究表明,配方D为本试验最适培养基配方,玉米芯60%、硬杂木屑18%、麸皮20%、石膏1%、糖1%。发菌期间,在配方D中的染菌率低,仅为3%,比其他配方低1%～4%;菌丝生长茁壮、浓密,45d满袋,比其他配方提早3～20d;平均产量0.245Kg,生物转化率为81.78%,比其他配方高8～34%。
(7)采用常规营养分析法对不同培养料所获的子实体营养成分进行测定,结果表明:不同配方所获的榆耳子实体水分含量在90%左右,差异不明显;但可溶性蛋白和可溶性糖含量差异较大,配方D中子实体可溶性蛋白9.06mg/100g,比其他配方高约1～2 mg/100g,达到极显著水平;可溶性糖含量为11.96mg/g,比其他配方高约1.5～4 mg/g,也达到极显著水平。不同培养料上收获的子实体颜色也有所差异,添加玉米芯的培养基配方的子实体颜色较浅。
综上所述,本试验筛选出1个玉米芯培养料专用型榆耳新品种,提出了菌丝最适培养温度和最适培养基配方;明确了榆耳具有完整的胞外酶系,纤维素为其主要营养源,半纤维素次之,木质素较少等营养生理特性。
Gloeostereum incamatum S.Ito et Imai is well-known as edible and medical fungi,eaten like sea cucumber,which has rich nourishment and the medicinal value for treatment of stomach disease.Recently, Gloeostereum incamatum has been artificially cultured initially and successfully in China,it has become one of the most important edible fungi in Northeast.But,quality and quantity is not stable due to the directly using wild species and lacking cultivation management experiment.Research on breeding of new variety and cultivation physiology of Gloeostereum incamatum should be developed immediately so as to keep the leading position of culturing management in China.In this paper,three excellent strains of C-1,C-2 and C-3 which output and quality were superior to their parents were selected from many strains and new species of Gloeostereum incamatum which cultivated with corncob culture materials was selected,Optimization of the compost formula,aimed to establish the basis of breeding and cultivation physiology,offer excellent new variety and corresponding cultivation technology in the corn productive region(northeast of China).The experimental results are as follows:
(1) According to antagonism identification,the tested strains were three different physiological characteristic ones which produced possession of specific antagonism line.
(2) The result of mycelium cultural character study showed:the temperature that three tested strains growing was between 10℃～35℃,the most suitable temperature was 25℃.Combining with growth speed, growth time and mycelium growth status showed that C-1 was the best strain.
(3) The result showed:the optimal adaptability to corncob medium was the strain of C-1.In mycelium growing stage,the contamination of C-1 was only 3%that was lower than the other strains 3%～5%.The mycelium was healthier and stronger,it grew faster and the cultivation bag was full of it in 50d which was earlier than the others for 3-7d.In fruit bodies budding period,the appearing bud time of C-1 was shorter than the other strains for 7～8d.Biological conversion ratio of fruit bodies can reach 66.5%which was 15%higher than the others.The fruit bodies were obviously excelled other strains in big size,thick,colorful(pink) and shape-like flower.
(4) The study on extracellular enzyme activity showed:it had complete extracellular enzyme systems, which could make lignocelluloses degradation completely in medium.Extracellular enzyme activity excreted in different medium formula behaved a certain disparity in each growing stage of Gloeostereum incamatum, but the general tendency of enzyme activity was essential same.The cellulose enzyme system had clearly complementary relationship and the activities of hemicellulase were stronger at the reproductive phase.The activity of lignocellulose enzyme system was higher at the mycelial growth stage and then decreased.
(5) The nutrition utilization regularity study showed:degradation ratios of all kinds of nutrition in each growth phase were different,the mostly utilization of nonlignocellulose and lignocellulose were mainly in the stage of mycelium growth;Form the change of amylase activity,starch materials were the main nutrition at the earlier stage,the main utilization of lignin was at the stage of mycelial growth;Hemicellulose was utilized mainly in the reproductive growth phase;But the utilization of cellulose was prolonged the whole growth phase.Finally,the average decrement of cellose was 51.39%,the hemicellulose was 32.22%and the lignin was 29.16%.Therefore,the test showed that cellulose was the major nutrition source of Gloeostereum incamatum,secondly was hemicellulose,lignin was fewer relatively.
(6) Through the study on mycelium growth condition,yield and biological conversion ratio in different medium,we could find that formula D was the most optimal medium which was composed of 60%corncob, 18%saw dust,20%wheat bran,1%calcium sulfate and 1%sucrose.During the period of culturing mycelium, the contamination ratio of formula D was only 3%which was lower than the other formulas by 1%～4%; mycelium was healthier and stronger and growing faster,filling bag in 45d which earlier 3～20d than the others;average yield was 0.245kg,biological conversion was 81.78%which was 8～34%higher than the other formulas.
(7) The nutrition analysis of fruit bodies which were obtained from different medium formulas showed: moisture content of fruit bodies were about 90%,the difference was not significance.But the content of soluble sugar and protein was in significant difference,the soluble protein of formula D was 9.06mg/100g which higher than the others for about 1～2mg/100g,getting a very significant level.The soluble sugar content was 11.96mg/g,which was higher than the others for about 1.5～4mg/g,reached extremely significant level. The color of fruit bodies which obtained from different medium formulas were different,the color of fruit body which obtained from corncob medium was tint.
Consequently,this experiment screened a best strain which was suitable to corncob medium cultivation and advanced the optimal temperature and formula of mycelium.Studying the change pattern of extracellular enzyme activities and cellulose was the major nutrition source of Gloeostereum incamatum,secondly was hemicellulose,lignin was fewer relatively.
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