浙江农业学报
浙江農業學報
절강농업학보
ACTA AGRICULTURAE ZHEJIANGENSIS
2014年
4期
941-947
,共7页
松茸%原生质体%释放方式%正交设计%再生率
鬆茸%原生質體%釋放方式%正交設計%再生率
송용%원생질체%석방방식%정교설계%재생솔
Tricholoma matsutake%protoplast%release modes%orthogonal experiment design%regeneration rate
借助正交设计法对松茸原生质体制备与再生进行研究，结果表明，松茸原生质体释放方式主要有4种，分别为顶端释放、侧位释放、菌丝段端位释放及原位释放。原生质体最佳制备体系是：菌龄2 d，以0.6 mol· L-1的KCl为渗透压稳定剂，10 L＋10 S＋10 C的酶系统34℃酶解2 h，最终制备率达到2.75×107个· mL-1；最佳再生体系为：菌龄2 d，以MgSO4·7H2 O为渗稳剂，10 L＋10 S的酶液30℃酶解4 h，在以蔗糖为渗稳剂的再生培养基上再生率最高，可达41.6％。
藉助正交設計法對鬆茸原生質體製備與再生進行研究，結果錶明，鬆茸原生質體釋放方式主要有4種，分彆為頂耑釋放、側位釋放、菌絲段耑位釋放及原位釋放。原生質體最佳製備體繫是：菌齡2 d，以0.6 mol· L-1的KCl為滲透壓穩定劑，10 L＋10 S＋10 C的酶繫統34℃酶解2 h，最終製備率達到2.75×107箇· mL-1；最佳再生體繫為：菌齡2 d，以MgSO4·7H2 O為滲穩劑，10 L＋10 S的酶液30℃酶解4 h，在以蔗糖為滲穩劑的再生培養基上再生率最高，可達41.6％。
차조정교설계법대송용원생질체제비여재생진행연구，결과표명，송용원생질체석방방식주요유4충，분별위정단석방、측위석방、균사단단위석방급원위석방。원생질체최가제비체계시：균령2 d，이0.6 mol· L-1적KCl위삼투압은정제，10 L＋10 S＋10 C적매계통34℃매해2 h，최종제비솔체도2.75×107개· mL-1；최가재생체계위：균령2 d，이MgSO4·7H2 O위삼은제，10 L＋10 S적매액30℃매해4 h，재이자당위삼은제적재생배양기상재생솔최고，가체41.6％。
This study aimed to explore the protoplast preparation and regeneration conditions of Tricholoma matsutake by orthogonal experiment .The results showed that there were four release modes of T.matsutake protoplasts:release from top,release from the side,hyphal fragment end release and release in situ .The optimal protoplast preparation condition was selecting mycelium cultured in liquid medium for 2 d, for enzymolysis in mixed enzyme solution containing 1% cellulose , 1% snailase and 1% lywallzyme at 34℃ for 2 h, with 0.6 mol· L-1 KCl as osmotic stabilizer,and the obtained preparation rate reached 2.75 ×107protoplasts· mL-1.The optimal regeneration condition was using mycelium cultured in liquid medium for 2 d,for enzymolysis in enzyme solution containing 1%lywallzyme and 1% snailase at 30℃ for 4 h,with 0.6 mol· L-1 MgSO4· 7H2O as osmotic stabilizer,and the regeneration rate reached 41.6%.
