摘要
适合番茄叶肉原生质体培养的材料是３～４周龄无菌苗顶端开展的叶片，在２７℃下，采用１０ｇ／Ｌ纤维素酶“Ｏｎｏｚａｋａ”Ｒ－１０、５ｇ／Ｌ果胶酶Ｐｅｃｔｉｎａｓｅ、５ｍｍｏｌ／ＬＭＥＳ、９０ｇ／Ｌ甘露糖醇、ＣＰＷ盐的酶液游离撕去下表皮的嫩叶，１４～１６ｈ，效果最好。研究表明，“红玫瑰”番茄和Ｌ．ｅｓｃ．×（Ｌ．ｅｓｃ．×Ｓ．ｌｙ．，Ｆ１），ＢＣ１Ｆ１的酶解效果最好，原生质体的产量和活力都最高；秘鲁番茄和Ｌ．ｅｓｃ．×Ｓ．ｌｙ．，Ｆ１次之；而潘那利番茄酶解效果最差，产量依次为：４８３×１０６、４５５×１０６、４３７×１０５、２１２×１０５和３０３×１０４个／ｇ。试验中，对多种培养基及培养方法进行对比研究，结果表明，Ｄ２培养基液体浅层法培养效果最好，ＴＭ－２液体浅层培养和Ｍ８Ｅ琼脂包埋漂浮法次之。在所采用的５种试材中，从“红玫瑰”番茄和Ｌ．ｅｓｃ．×（Ｌ．ｅｓｃ．×Ｓ．ｌｙ．，Ｆ１），ＢＣ１Ｆ１的叶肉原生质体获得了愈伤组织。
uitable materials for tomato protoplast isolation and culture were leaves from 3～4 week-old axenic seedlings grown in agar(in test tube). The young leaves torn lower epidermis were incubated in an enzyme solution consisting of 10 g/L “Onozaka” R-10 cellulase,5 g/L pectinase, 5 mmol/L MES and 90 g/L mannitol in CPW salts for 14～16 h at 27℃. The mesophyll protoplast yields were 4.83×106,4.55×106,4.37×105,2.12×105 and 3.03×104 protoplasts/gram fresh leaves respectively for L.esc. cv. Red Rose, L.esc. × (L.esc. × S.ly.,F1),BC1F1 ,L.peruvianum, L. esc × S.ly., F1 and S. pennellii, all with viability of more than 80% but less than 90% .The D2 liquid culture was the most suitable in several medium and culture methods With this method, micro calli had been obtained from the mesophyll protoplasts of L.esc. cv,Red Rose and L. esc. × (L.esc.× S.ly.,F1), BC1F1.Transferred to agar medium for enhancement after the calli grew to 1 to 2 mm in diameter, it ceased to grow and came to brown.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
1997年第4期36-41,共6页
Journal of South China Agricultural University
关键词
番茄
叶肉原生质体
组织培养
育种
Lycopersicon esculentum(L.esc.)
wild tomato apecies
mesophyll protoplasts
