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Induction and early identification of tetraploid black locust by hypocotyl in vitro
In Vitro Cellular & Developmental Biology - Plant ( IF 2.6 ) Pub Date : 2021-01-14 , DOI: 10.1007/s11627-020-10133-5
Xiuyu Li , Zijie Zhang , Yunhui Ren , Yue Feng , Qi Guo , Li Dong , Yuhan Sun , Yun Li

Tetraploid black locust (Robinia pseudoacacia L.) has advantages over diploid varieties in growth and stress resistance. Previous studies had shown that tetraploid black locust can be induced only by treating shoot tips of the germinating seeds with colchicine. In order to establish a simpler and more efficient tetraploid induction method in black locust, we induced tetraploid plants by treating the hypocotyls from in vitro–grown seedlings with different concentrations of colchicine, and then provided a method for tetraploid identification based on stomatal size and density in this study. The highest induction efficiency was 53.33% by treating hypocotyl explants with 70 mg·L−1 colchicine for 2 d. Tetraploid black locust was significantly different from diploid in morphological characteristics. Leaf area, leaf thickness, chlorophyll content, stomatal length, and stomatal width were greater in tetraploid plants than in diploid plants, whereas the opposite was true for leaf aspect ratio, stomatal index, and stomatal density. The differences in stomatal size and density between diploid and tetraploid were nearly two times; therefore, size and density of stomata can be used as a stable and distinct marker to rapidly screen tetraploids. The results of this study suggested that the hypocotyl was an ideal material to induce tetraploid black locust in vitro and confirmed that stomatal size and density could be an effective and reliable marker to identify ploidy level in black locust.



中文翻译:

下胚轴对四倍体黑刺槐的诱导及早期鉴定

四倍体黑刺槐(Robinia pseudoacacia L.)在生长和抗逆性方面优于二倍体变种。先前的研究表明,只有用秋水仙碱处理发芽种子的梢部才能诱导四倍体黑刺槐。为了建立一种简单,高效的刺槐四倍体诱导方法,我们用不同浓度的秋水仙碱处理体外生长的幼苗的下胚轴,从而诱导了四倍体植物,然后提供了一种基于气孔大小和密度的四倍体鉴定方法。在这个研究中。用70 mg·L -1处理下胚轴外植体时,最高诱导效率为53.33%。秋水仙碱2 d。四倍体黑刺槐的形态特征与二倍体显着不同。四倍体植物的叶面积,叶厚度,叶绿素含量,气孔长度和气孔宽度大于二倍体植物,而叶的长宽比,气孔指数和气孔密度则相反。二倍体和四倍体之间的气孔大小和密度差异几乎是两倍。因此,气孔的大小和密度可用作快速筛选四倍体的稳定且独特的标记。这项研究的结果表明,下胚轴是体外诱导四倍体黑刺槐的理想材料,并证实气孔大小和密度可能是鉴定黑刺槐倍性水平的有效和可靠的标记。

更新日期:2021-01-14
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