旱芹人工种子内胚状体的优化

THE OPTIMIZATION OF EMBRYOID FOR ARTIFICIAL SEED OFApium graveolens

分别以旱芹（ＡＰｉｕｍｇｒａｖｅｏｌｅｎｓ）的天然种子、子叶、真叶等为外植体，在ＭＳ＋２，４－Ｄｌｍｇ／Ｌ＋ＫＴ０．５ｍｇ／Ｌ的培养基上诱导产生胚状体包埋成的人工种子，可获得最高的转株率和最低的畸形率。四种愈伤组织的ＳＤＳ－ＰＡＧＥ电泳结果表明，子叶愈伤组织的蛋白成分比其它三种愈伤组织缺少８６ｋＤ、６３ｋＤ、１７ｋＤ、１５ｋＤ、１３ｋＤ的蛋白，这可能决定了子叶胚状体的高质量。另外，用ＭＳ＋ＡＢＡ１ｍｇ／Ｌ处理子叶愈伤组织，可使胚状体的蛋白含量从１．３ｍｇ／ｇ（鲜重）提高到２．０ｍｇ／ｇ，从而大大提高了人工种子冷藏后的转株率。所以，胚状体的优化是必要的，也是可行的。

Calli were induced from different.explants such as natural seed, leaf, cotyledon of Apium graveolens on MS media supplemented with lmg/L 2,4-D and 0. 5mg/L KT.The somatic embryos were produced by inoculating the callus from cotyledon into hormonefree MS medium, and were constructed into artificial seeds. This kind of artificial seeds obtained the highest conversion rate and the lowest deformation rate among those kinds of artificial seeds from different explants. The SDS-PAGE electrophoresis result indicated that the protein ingredient of callus from cotyledon was less than other kinds of calli. Maybe, the lack of 86kD, 63kD, 17kD, 15kD, 13kD resulted in the high quality of artificial seeds from cotyledon. In addition, when ABA was applied at the preferable concentration ling/L to treat the callus from cotyledon, the concentration of protein was increased from 1. 3mg/g (fresh weight) to 2. 9mg/g. The artificial seeds constructed from ABA-treated embryoids gained 56. 1 % conversion rate after 30 days low temperature storage. These results indicated the suitability and necessity of optimization of embryoids.