短季大豆基因型增殖胚胎发生和基因枪转化的基因型筛选

Genotype screening for proliferative embryogenesis and biolistic transformation of short-season soybean genotypes.

作者信息

Simmonds D H, Donaldson P A

机构信息

Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Bldg. 21, Central Experimental Farm, Ottawa, Ontario, Canada K1A OC6 e-mail:

出版信息

Plant Cell Rep. 2000 Apr;19(5):485-490. doi: 10.1007/s002990050760.

DOI:10.1007/s002990050760

PMID:30754887

Abstract

Eighteen of 20 short-season soybean (Glycine max (L.) Merrill) genotypes (maturity group 0 and 00) screened for proliferative embryogenic capacity formed secondary globular embryos, at rates of 1-70% of cultured immature cotyledons. Five genotypes produced embryogenic cultures which were proliferative for at least 6 months. Proliferative embryogenic cultures of AC Colibri and X2650-7-2-3 were bombarded using a Bio-Rad PDS-1000/He particle gun. Co-bombardments with plasmid pairs pHygr (encoding a type IV aminoglycoside phosphotransferase;aphIV) and pRD300pat (encoding a phosphinothricin N-acetyltransferase;pat) or pRD300pat and pFF19G (β-glucuronidase;uidA or gus) resulted, respectively, in 12 hygromycin-selected lines with multiple insertions of aphIV and pat, and two l-phosphinothricin-selected lines plus three β-glucuronidase-positive lines recovered without selection. Although fertile plants were recovered from young proliferative cultures, transgenic plants, which were derived from cultures 12-14 months of age, were sterile.

摘要

在筛选具有增殖胚性能力的20个短季大豆（Glycine max (L.) Merrill）基因型（成熟组0和00）中，有18个形成了次生球形胚，占培养未成熟子叶的1%-70%。5个基因型产生了胚性培养物，其增殖至少持续6个月。使用Bio-Rad PDS-1000/He粒子枪对AC Colibri和X2650-7-2-3的增殖胚性培养物进行轰击。用质粒对pHygr（编码IV型氨基糖苷磷酸转移酶；aphIV）和pRD300pat（编码膦丝菌素N-乙酰转移酶；pat）或pRD300pat和pFF19G（β-葡萄糖醛酸酶；uidA或gus）共轰击，分别得到了12个具有多个aphIV和pat插入的潮霉素筛选系，以及2个膦丝菌素筛选系和3个未经过筛选回收的β-葡萄糖醛酸酶阳性系。虽然从年轻的增殖培养物中获得了可育植株，但来自12-14个月龄培养物的转基因植株是不育的。