桦纤孔菌种质资源评价及矿质元素富集的生理效应研究

发布时间：2018-04-01 00:19

本文选题：桦纤孔菌　切入点：生物学性状　出处：《沈阳农业大学》2015年博士论文

【摘要】：桦纤孔菌Inonotus obliquus (Ach.ex Pers.) Pilat,是一种十分珍稀的食药用菌,非常具开发的潜力。药用部分为菌核,可治疗多种疑难疾病,例如高血压、癌症、糖尿病等。其中,对糖尿病的有效率可高达93%,同时具有一定抗肿瘤活性。随着人民生活水平的提高,其原料的需求量也逐年增加,但由于桦纤孔菌菌株间的生物学性状和品质差异较大,严重制约了桦纤孔菌的发展。为了更好地发挥桦纤孔菌的资源优势,本研究对不同桦纤孔菌进行了资源评价及矿质元素富集的作用研究,主要研究结果如下：1.应用SRAP、ITS分子标记技术对22个桦纤孔菌菌株进行资源鉴定,结果表明：HLJ01, JL01, JL02, JL03, JL04, JL05, Z2, CX02, FH, NBRC9788, MAFF420239, MAFF420102, MAFF420243 和 MAFF420279的亲缘关系非常近。他们中有七个菌株来自中国,一个来自朝鲜,一个来自芬兰,一个来自美国,四个来自日本。菌株MAFF420256和HE在两种标记分析中的结果不一致。MAFF410028和NBRC8681在ITS分析中表现出了很近的亲缘关系,但SRAP分析将MAFF410028单独划为一类。MAFF420348 和 MAFF420165在两种分析中均表现出很近的亲缘关系。MAFF420198在两种分析方法中也表现出了不同结果。两种分子标记表明来自中国的菌株亲缘关系较近。来自日本的菌株亲缘关系较复杂。这表明亲缘关系与地理纬度关系密切。日本的地理纬度差异较大,个别菌株亲缘关系较远。中国的菌株及朝鲜的菌株地理纬度差异较小,表现出了较近的亲缘关系。2.本文对桦纤孔菌的菌丝进行了生物学性状研究。包括桦纤孔菌菌株的菌丝显微结构、拮抗性、菌核栽培、不同生长调节剂对胞外酶活性影响等方面。结果表明：菌株JL01菌丝直径最大,为4.20μm,与MAFF420101菌丝直径较为接近,显著优于其他菌株；菌株FH拮抗青霉的能力最强,菌丝长势最好,菌圈直径最大可达53.27mm；FH菌株生物学效率最高,菌核干重达25.81g/袋,生物学效率达26.12%；当添加2,4-D时胞外纤维素酶FP酶与CMC酶活性均最高,可达8.12 U/g和13.11 U/g, KT处理的淀粉酶活性最高；6-BA处理的胞外蛋白酶活性在各个时期均显著优于其他处理,在菌核成熟期可以达到107 U/g。综上所述,JLO1, MAFF420 102, FH, HE适合人工栽培,NBRC8681,MAFF420256, NBRC9788, MAFF420239, MAFF410028菌株性状较差,均不适合人工栽培。3.通过对桦纤孔菌有效化学成分、矿质元素、营养成分含量的分析,结果显示：菌株JL01白桦脂醇含量较高,为23.7mg/g。 FH 次之。 MAFF420165、MAFF420348白桦脂醇含量均较低,不适合生产白桦脂醇。菌株FH多糖含量最高,可达9.7%,显著优于其他菌株,是开发多糖产品的优良菌株；菌株NBRC9788钙含量最高,达3.49 mg/g,显著优于其他菌株。菌株FH其次；MAFF420165的菌株含磷量最高,高达210.1μt/g,显著高于其他菌株的磷含量；来自芬兰的菌株FH游离水含量最高,达到77.21%,来自吉林的菌株JL01结合水含量最高,达10.74%；菌株FH粗脂肪、粗蛋白含量均最高,分别为1.93%和20.23%；粗灰分含量最高者MAFF420165,达到8.76%；粗纤维含量较高的是来自中国的JL01,高达45.09%。综上所述,菌株FH, JL01, MAFF420165有效化学成分含量较高,适合开发使用。菌株NBRC8681、MAFF420101, MAFF420348, MAFF420239有效化学成分含量较低。4.以桦纤孔菌为材料,研究了不同矿质元素钙,铁,锌在其菌丝体中的富集及生长的影响。结果表明：在培养基中添加1000mg/L的Ca (NO3)2能明显促进菌丝体生长和产量提高,钙的富集率可高达0.07%。桦纤孔菌菌丝生长最适宜的铁源是Fe2 (SO4) 3,添加浓度为300 mg/L,富铁率最高为0.29%,优于其他处理。添加ZnSO4浓度为175mg/L的处理菌丝长势最强,富集率最高可达0.57%；富集300 mg/L的Fe2(SO4)3处理对桦纤孔菌抗氧化活性整体效果较好,富集矿质元素锌对还原力提高和富集矿质元素钙对抗卵黄脂质清除率提高作用效果最为明显；富集1000 mg/L的Ca(NO3)2处理,桦纤孔菌菌株粗多糖,蔗糖,果糖的含量均最高,分别为15.63%,14.76%,13.08%；富集矿质元素Fe2(SO4)3浓度为300 mg/L的处理氨基酸、粗脂肪、粗灰分含量均最高。富集矿质元素ZnSO4浓度为175 mg/L处理滤纸纤维素酶、蛋白酶活性均最高。富集Ca(NO3)2浓度为1000 mg/L处理羧甲基纤维素酶、淀粉酶活性最高。
[Abstract]:Inonotus intermedius ( AchE . ex Maxim . ) The results are as follows : 1 . There are seven strains from China , one from North Korea , one from Finland , one from the United States and four from Japan . Compared with other strains , the diameter of the mycelium of MAFF420101 was close to that of MAFF420101 .
The strain FH antagonistic Penicillium was the most powerful , the length of the mycelium was the best , the diameter of the bacteria circle was up to 53.27mm ;
The biological efficiency of FH strains was the highest , the dry weight of sclerotium reached 25.81 g / bag , and the biological efficiency reached 26.12 % .
When 2,4 - D was added , the activity of extracellular cellulase FP and CMC was highest , which could reach 8.12 U / g and 13.11 U / g , KT - treated amylase had the highest activity .
In conclusion , JLO1 , MAFF420 102 , FH , HE were suitable for artificial cultivation , NBRC8681 , MAFF420256 , NBRC9788 , MAFF420239 and MAFF410028 were not suitable for artificial cultivation . FH the second time . The content of MAFF420165 , MAFF420348 and betulin was lower , which was not suitable for the production of betulin . The highest content of polysaccharide in strain FH was 9.7 % , which was superior to other strains , and it was an excellent strain for development of polysaccharide products .
The highest calcium content of strain NBRC9788 was 3.49 mg / g , which was superior to other strains .
The strain of MAFF420165 had the highest phosphorus content , up to 210.1渭t / g , which was significantly higher than that of other strains .
The free water content of strain FH from Finland was the highest , reaching 77.21 % , and the water content of JL01 strain JL01 from Jilin was up to 10.74 % .
The crude protein content of strain FH was 1.93 % and 20.23 % , respectively .
MAFF420165 , the highest ash content , reached 8.76 % ;
In conclusion , the content of effective components of strain FH , JL01 and MAFF420165 was high , which was suitable for the development and use . The results showed that the content of effective components of strain NBRC8681 , MAFF420101 , MAFF420348 and MAFF420239 was lower .
The concentration of Fe 2 ( SO4 ) 3 enriched in 300 mg / L has a good effect on anti - oxidation activity of Fuscoporia Obliques , and the effect of enriching mineral element zinc on reducing power and enriching mineral element calcium against yolk lipid clearance is most obvious .
The content of crude polysaccharide , sucrose and fructose was highest at 1000 mg / L , 15.63 % , 14.76 % and 13.08 % respectively .
The concentration of Fe 2 ( SO4 ) 3 was 300 mg / L , and the content of crude fat and crude ash was the highest . The concentration of enrichment mineral element ZnSO4 was 175 mg / L , and the activity of protease was the highest . The concentration of Ca ( NO3 ) 2 was 1000 mg / L , and the activity of amylase was the highest .

【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S567.39

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