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Improvement of Callus Induction Efficiency in Anther Culture of Autotetraploid Rice

提高水稻同源四倍体花药培养愈伤诱导率的研究


以同源四倍体水稻原种零轮、02428、培矮64、轮回422及其成对二倍体为实验材料,分别以琼脂浓度(0.5%、0.75%和1%)、碳源种类(蔗糖和麦芽糖)与用量(60 g L-1和80 g L-1)、不同激动素(KT和Zip)以及不同基本诱导培养基(MS、N6、M8、NB)等因素为变量,共设计26种诱导培养基(2种对照),进行了花药愈伤诱导实验,旨在筛选出愈伤诱导率的培养基优化组合。结果表明,琼脂浓度0.75%和60 g L-1蔗糖组合对提高水稻花药诱导率作用较好, N6培养基及其衍生NB培养基对提高四倍体水稻特别是粳稻花药诱导率有着较好作用。激动素的加入,能显著提高愈伤诱导率,最高可达到84%。在供试的26种培养基中,有6种诱导培养基得到较高的四倍体粳稻愈伤诱导率,有2种诱导培养基得到较高的四倍体籼稻愈伤诱导率,在8种较好的诱导培养基中有3种不但有较高的愈伤诱导率而且具有较高的绿苗率。

Improving anther culture inductivity is important for improvements of plant differentiation rate and plant production rate in rice. Four pedigree seed varieties of autotetraploid rice, Linglun, 02428, Pei’ai 64 and Lunhui 422 and the corresponding diploid plants were used in the present work of anther culture. 26 kinds of induction medium were designed on the factors including agar concentration (0.5%, 0.75% and 1%), variety (sucrose and maltose) and concentration (60 and 80 g L-1) of carbon source, different varieties of basic media (MS, N6, M8, NB) and hormone (KT and Zip) to determine appropriate combination on callus induction. The results indicated that the inductivity of 60 g L-1 (18.7%) is notably better than that of 80 g/L (8.7%) when using sucrose as carbon source. Meanwhile, using 0.75% of agar obtained the highest inductivity which is 20.8% among twelve media combination, and the average induction rate of autotetraploid japonica (16.4%) is higher than that of autotetraploid indica (7.3%). The N6 and NB media were better than MS and M8 for improving callus inductivity in tetraploid rice, especially in Japonica rice, whether sucrose or maltose was used as carbon source. The media with supplement of KT and Zip could induce more calli conspicuously, and the highest inductivity was 84%. It was also observed that the calli induced by the addition of KT were of high quality to produce seedlings. Among total 26 kinds of media, 6 types led to higher inductivity in japonica rice whereas 2 types in indica rice. Furthermore, there were 3 types media produced not only higher callus inductivity but also higher plant production rate than others. In conclusion, the induction efficiency of anther callus for one certain genotype is remarkably different with various media, which implies that the genotype is of selection for the induction media.


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