XIN Shiji,LIU Jingwen,YANG Fan,et al.Breeding High-yield Strain of Pleurotus nebrodensis by ARTP Mutagenesis Technology[J].Northern Horticulture,2023,(23):115-121.[doi:10.11937/bfyy.20232265]
利用ARTP诱变技术选育白灵菇高产菌株
- Title:
- Breeding High-yield Strain of Pleurotus nebrodensis by ARTP Mutagenesis Technology
- 文章编号:
- 1001-0009(2023)23-0115-07
- 关键词:
- 白灵菇; 原生质体; 常压室温等离子体（ARTP）诱变
- 分类号:
- S 646.9
- 文献标志码:
- A
- 摘要:
- 以白灵菇“中农1号”为试材，研究了白灵菇“中农1号”原生质体制备条件；以不做处理的“中农1号”为CK，首次采用常压室温等离子体（atmospheric room temperature plasma，ARTP）诱变技术，研究了其对菌株诱变后子实体产量的影响，以期获取相比CK菌株产量更高的优良菌株。结果表明：通过单因素试验得到白灵菇“中农1号”原生质体的最佳制备条件为酶解温度28 ℃、酶解时间3 h、溶壁酶浓度为20 mg·mL-1、渗稳剂使用0.6 mmol·L-1 KCl。ARTP诱变最佳时间为60 s，致死率为94%时正突变率最为稳定。通过平板筛选试验初筛结果显示，生长速度快、产漆酶、纤维素酶能力强的菌株有4株，进一步通过了LBL脱色试验复筛，筛选出结实性好的4株菌株ZN-37、ZN-20、ZN-39、ZN-19。通过测定白灵菇栽培产量确定最终的高产菌株为ZN-19，总产量高达966.95 g，相比CK菌株总产量多37%。
- Abstract:
- The protoplast preparation conditions of Pleurotus nebrodensis ‘Zhongnong No.1’ were studied by using Pleurotus nebrodensis ‘Zhongnong No.1’ as test material.The ‘Zhongnong No.1’ without treatment was used as CK,and the atmospheric room temperature plasma ( ARTP ) mutagenesis technique was used for the first time to study its effect on the yield of fruiting bodies after mutagenesis,in order to obtain superior strains with higher yield than CK.The results showed that the optimum preparation conditions of protoplasts of Pleurotus nebrodensis ‘Zhongnong No.1’ obtained by single factor test were as follows，enzymolysis temperature 28 ℃,enzymolysis time 3 hours,lywallzyme concentration 20 mg · mL-1,osmotic stabilizer 0.6 mmol · L-1 KCl.The optimal time of ARTP mutagenesis was 60 seconds,and the positive mutation rate was the most stable when the lethal rate was 94%.The preliminary screening results of the plate screening test showed that there were 4 strains with fast growth rate,strong laccase and cellulase production ability.Further,the LBL decolorization test was rescreened,and 4 strains ZN-37,ZN-20,ZN-39 and ZN-19 with good seed setting were screened out.By measuring the cultivation yield of Pleurotus nebrodensis,the final high-yield strain was determined to be ZN-19,with a total yield of 966.95 g,which was 37 % more than the total yield of the CK group.
参考文献/References:
［1］DING J H，HAN X N，LIU S W，et al.Sesquiterpenoids from the edible mushroom Pleurotus nebrodensis［J］.Chemistry of Natural Compounds，2022，58(1)：37-39.［2］KIM I H，LEE J H.Skin whitening and anti-wrinkle effects of Pleurotus nebrodensis extracts［J］.KSBB Journal，2021，36(3)：209-215.［3］ASLAM S，BILAL M.Pleurotus-derived laccases，immobilization，and bioremediation applications［M］//MULLA S I,BHARAGAVA R N.Enzymes for Pollutant Degradation.Singapore：Springer Nature Singapore，2022：49-69.［4］DOS REIS E E，SCHENKEL P C，CAMASSOLA M.Effects of bioactive compounds from Pleurotus mushrooms on COVID-19 risk factors associated with the cardiovascular system［J］.Journal of Integrative Medicine，2022,20(5):385-395.［5］MISHRA V，TOMAR S，YADAV P，et al.Promising anticancer activity of polysaccharides and other macromolecules derived from oyster mushroom (Pleurotus sp.)：An updated review［J］.International Journal of Biological Macromolecules，2021，182：1628-1637.［6］GOLAK-SIWULSKA I，KAU〖AKZ·〗EWICZ A，SPI〖AKZ·〗EWSKI T，et al.Bioactive compounds and medicinal properties of Oyster mushrooms (Pleurotus sp.)［J］.Folia Horticulturae，2018，30(2)：191-201.［7］CHU L，YANG L，LIN L，et al.Chemical composition，antioxidant activities of polysaccharide from pine needle (Pinus massoniana) and hypolipidemic effect in high-fat diet-induced mice［J］.International Journal of Biological Macromolecules，2019，125：445-452.［8］曹瑶,闻绍锋,刘书畅,等.白灵菇研究进展综述［J］.食药用菌,2019,27(3):169-173.［9］常存,段楠,杨雪冰.白灵菇的营养成分测定及保健功能研究［J］.黑龙江科学,2019,10(14):48-49.［10］朱慧霞,方桢,鲁旭峰,等.常压室温等离子体诱变选育木聚糖酶高产菌及其酶学性质研究［J］.中国酿造,2019,38(7):31-36.［11］金鸣,王威,张雪洪.高产洛蒙真菌素的洛蒙德链霉菌的高通量筛选方法建立与复合育种［J］.微生物学通报，2020，47(1):200-209.［12］宋甜甜,吴迪,张赫男,等.ARTP诱变猴头菌株的发酵菌丝体多糖理化性质及体外免疫活性［J］.菌物学报,2018,37(6):794-804.［13］任林英,张祝兰,唐文力,等.常压室温等离子体诱变选育替考拉宁高产菌株［J］.中国抗生素杂志,2018,43(1):46-50.［14］YUAN L I，HENAN Z，YI T A N，et al.Screening of a high polysaccharide content Ganoderma lucidum strain by ARTP［J］.Acta Edulis Fungi，2021，28(2)：36-44.［15］SUN S,LI X,RUAN L,et al.A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa based on ligninolytic enzymes［J］.World Journal of Microbiology and Biotechnology，2014，30(7):2005-2013.［16］吴晓华.真姬菇与金针菇原生质体融合育种研究［D］.福州:福建农林大学,2011.［17］王伟,郑大浩,杨超博,等.高效纤维素分解菌的分离及秸秆降解生物效应［J］.中国农业科技导报,2019,21(8):36-46.［18］刘洋,陈辉,冯志勇,等.LBL评价与真姬菇表观农艺性状相关性分析［J］.上海农业学报,2011,27(2):116-120.［19］娄海伟,余颖豪,林俊芳,等.基于蛹虫草单核芽生孢子的原生质体制备条件的优化［J］.中国食品学报,2020,20(6):129-136.［20］何小慧.杏鲍菇原生质体再生变异菌株的筛选［D］.福州:福建师范大学,2014.［21］陈力力,张倩云,甘文娟,等.金针菇菌种诱变选育的研究［J］.生物技术,2011,21(2):67-69.［22］李慧玲,刘永梅.诱变选育高产DHA裂殖壶菌突变株［J］.食品科技，2015，40(9):12-16.［23］陈琼华,周玉萍,毕凡星,等.真菌漆酶高产菌株的筛选［J］.广州大学学报(自然科学版),2009,8(5):53-57.［24］李莎,刘宇,马康,等.白灵菇菌种退化的快速鉴别方法［J］.生物技术,2014,24(6):66-68.［25］原慧,王勇,周继阳.新疆阿魏菇高效原生质体分离与再生体系的建立［J］.食用菌,2012,34(2):12-14.［26］刘文芳,郭成金.白灵菇原生质体制备与再生研究［J］.天津师范大学学报(自然科学版),2012,32(1):88-92.［27］陈辉,刘洋,赵楚楚,等.真姬菇菌种质量评价方法的优化研究［J］.上海农业学报,2013,29(4):4-8.
相似文献/References:
[1]张金鹏,杨 晶,韩玉珠,等.植物体细胞杂交的研究进展[J].北方园艺,2014,38(01):192.
ZHANG Jin-peng,YANG Jing,HAN Yu-zhu,et al.Research Progress of Plant Somatic Hybridization[J].Northern Horticulture,2014,38(23):192.
[2]刘 宇,王兰青,王守现,等.棉柴栽培白灵菇试验[J].北方园艺,2013,37(17):148.
LIU Yu,WANG Lan-qing,WANG Shou-xian,et al.Utilization of Cotton Stem on Pleurotus nebrodensis Cultivation[J].Northern Horticulture,2013,37(23):148.
[3]邓红梅,马超,王颖.黄瓜幼苗子叶原生质体的分离纯化及融合条件优化研究[J].北方园艺,2013,37(11):6.
DENG Hong-mei,MA Chao,WANG Ying.Study on the Optimal of Separation,Purification and Fusion Conditions of Cucumber Seedlings Cotyledons Protoplast[J].Northern Horticulture,2013,37(23):6.
[4]王兰青,刘宇,王守现,等.工厂化栽培白灵菇配方筛选试验[J].北方园艺,2012,36(01):156.
WANG Lan-qing,LIU Yu.,WANG Shou-xian,et al.Screening of Culture Medium for Industrial Cultivation of Pleurotus nebrodensis[J].Northern Horticulture,2012,36(23):156.
[5]徐琴,王晓军,王卉,等.苘麻叶肉原生质体的分离和培养研究[J].北方园艺,2013,37(07):170.
XU Qin,WANG Xiao-jun,WANG Hui,et al.Study on Isolation and Culture of the Mesophyll Protoplasts from Abutilon avicennae[J].Northern Horticulture,2013,37(23):170.
[6]崔瑞峰,马瑞霞.白灵菇二潮菇最佳出菇方式研究[J].北方园艺,2013,37(16):178.
CUIRui-feng,MARui-xia.StudyontheBestSecondFruitingWaysof Pleurotusnebrodensis[J].Northern Horticulture,2013,37(23):178.
[7]王昱,王义,王康宇,等.灵芝原生质体的制备与再生研究[J].北方园艺,2013,37(16):184.
WANGYu,WANGYi,WANGKang-yu,et al.StudyonPreparationandRegenerationoftheProtoplastsfrom Ganodermalucidum[J].Northern Horticulture,2013,37(23):184.
[8]王谦,马旭光,刘敏,等.原生质体紫外诱变选育猴头菌株[J].北方园艺,2014,38(16):145.
WANG Qian,MA Xu-guang,LIU Min,et al.Selection for a Hericium erinaceus Mutant Strain in the Way of Protoplasts Being Exposed to the UV Radiation[J].Northern Horticulture,2014,38(23):145.
[9]王卉,宁慧霞,刘敏,等.莴苣原生质体的分离方法[J].北方园艺,2012,36(11):109.
WANGHui,NINGHui-xia,LIUMin,et al.StudyontheMethodofIsolationProtoplastsofLactucasativaL.[J].Northern Horticulture,2012,36(23):109.
[10]肖 望,涂红艳,邓崇会.红姜花胚性细胞悬浮体系的建立和原生质体培养[J].北方园艺,2015,39(05):104.[doi:10.11937/bfyy.201505032]
XIAO Wang,TU Hong-yan,DENG Chong-hui.Establishment of Embryogenic Cell Suspension and Protoplast Culture of Hedychium coccineum[J].Northern Horticulture,2015,39(23):104.[doi:10.11937/bfyy.201505032]
备注/Memo
第一作者简介：辛世纪（1999-），男，硕士研究生，研究方向为菌类作物栽培育种。E-mail：spdyxsj@163.com.责任作者：黄亮（1983-），男，博士，教授，现主要从事农业生物技术及菌类作物等研究工作。E-mail：huangliang@tjau.edu.cn.基金项目：中央引导地方科技发展专项资助项目（20ZYCGSN00330）；天津“131”创新性人才培养工程资助项目。收稿日期：2023-07-05