甜荞种子多倍体诱导及其C1代DNA与形态学鉴定

Polyploid induction of common buckwheat seeds and its C1 generation DNA and morphological identification

利用秋水仙素对甜荞种子多倍体诱导处理,探讨0.1%(A_(1))、0.2%(A_(2))两种浓度的秋水仙素和6(B_(1))、9(B_(2))、12(B_(3))、18(B_(4))、24 h(B_(5))浸泡处理时间下最优的诱导加倍组合,通过种子萌发后C1代植株DNA鉴定选育出多倍体甜荞植株,对其形态学特征、气孔和保卫细胞特征进行鉴定。结果表明,以A_(2)B_(3)处理组合的诱导率最高,为80%;其次是A_(1)B_(5)、A_(2)B_(5)的处理组合,诱导率分别为75%和50%。经流式细胞仪对C1群体DNA进行鉴定,从成苗的193个植株中共获得65株变异植株,其中36株为四倍体,29株为非整倍体。形态学鉴定发现,与二倍体对照荞麦相比,多倍体甜荞的株高降低、茎粗增加、分枝减少;根系粗壮、须根增多;叶片表面皱缩粗糙、叶色加深;花器官变大、颜色更加鲜艳;种子变大且种皮外表呈灰色花纹。叶片下表皮气孔面积变大,四倍体、非整倍体的长度分别增加了11.44、20.52μm,且单位面积气孔数目分别减少了9个、10个。

Using colchicine to induce polyploidy of common buckwheat seeds,two colchicine concentrations of 0.1%(A_(1))and 0.2%(A_(2))and five soaking periods of 6h(B_(1)),9h(B_(2)),12h(B_(3)),18h(B_(4)),and 24h(B_(5))were studied.Polyploid common buckwheat plants were selected by DNA identification of C1 generation plants after seed germination.The characteristics of morphology,stomata,and guard cells were identified.The results showed that the induction rate of A_(2)B_(3)treatment combination was the highest,which was 80%,followed by A_(1)B_(5)and A_(2)B_(5)treatment combination,and the induction rate was 75%and 50%,respectively.The DNA of C1 population was identified by flow cytometry.A total of 65 variant plants were obtained from 193 plantlets,of which 36 were tetraploid and 29 were aneuploid.Morphological identification showed that,compared with the diploid control,the plant height and branches of polyploid common buckwheat decreased and its stem diameter and fibrous root increased.Its root system was strong,leaf surface shrunk rough,and leaf color deepened.The polyploid common buckwheat floral organ became larger and brighter than that of the diploid control common buckwheat.Its seed became larger and the seed coat showed a gray pattern.The stomatal area of lower epidermis of leaves increased,and the length of tetraploid and aneuploid increased by 11.44、20.52μm.The number of stomata per unit area decreased by 9 and 10,respectively.