| 流式细胞术在测定竹类植物基因组大小中的应用 |
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| 引用本文： | 陈蓉芬,黄坚钦,陈荣,徐川梅.流式细胞术在测定竹类植物基因组大小中的应用[J].浙江农林大学学报,2021,38(1):103-111. |
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| 作者姓名： | 陈蓉芬 黄坚钦 陈荣 徐川梅 |
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| 作者单位： | 浙江农林大学 省部共建亚热带森林培育国家重点实验室，浙江 杭州 311300 |
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| 基金项目： | 浙江农林大学校课堂教学改革项目(KGZD18001);浙江省自然科学基金资助项目(LY13C160011)。 |
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| 摘 要： | 目的 分析竹类植物不同组织部位及不同处理方法对其基因组大小的影响，可提高植物基因组大小测定精度。 方法 以竹类植物叶片和笋为材料，以水稻Oryza sativa为参照，设置细胞核染色时间为1、3、5、7、9、12、18、24和30 min共9个梯度，利用流式细胞仪对不同组织部位及处理的基因组大小进行分析。 结果 ①对同一竹种而言，其叶片和笋流式峰形图相似，基因组大小仅存在微小差异，差值为0.04~0.20 pg。②不同竹种对染色时间要求不同，其中，唐竹Sinobambusa tootsik、花叶唐竹Sinobambusa tootisik f. albo-striata、平安竹Pseudosasa japonica var. tsutsumiana、曙筋矢竹Pseudosasa japonica f. akebono、美丽箬竹Indocalamus decorus、花叶赤竹Sasaella glabra f. albo-striata及红秆寒竹Chimonobambusa mamorea f. variegata染色1 min即达到最大荧光峰值，孝顺竹Bambusa multiplex和黄皮绿筋竹Phyllostachys sulphurea染色3 min达到最大峰值，茶竿竹Pseudosasa amabilis var. amabilis和柳叶细竹Thyrsostachy ssiamensis染色5 min达到最大峰值，仅黄皮刚竹Phyllostachys sulphurea(叶片)7 min达到最大峰值。③12个竹种的荧光强度在1~30 min内存在较大变化，除茶竿竹、柳叶细竹、孝顺竹叶片及唐竹笋外，其他竹种的荧光强度变化值均在5%以上，特别是平安竹和花叶赤竹变化更大，分别为12.93%和12.88%。④热带木本竹种孝顺竹和柳叶细竹基因组大小为(1.64±0.54)~(2.69±1.01) pg，其他10个温带竹种基因组大小为(3.76±1.51)~(5.73±1.85) pg。10个温带竹种中，刚竹属Phyllostachys竹种基因组较小，大小为(3.76±1.51)~(3.91±0.95) pg，其他竹属的一些竹种基因组较大，大小为(4.82±0.54)~(5.73±1.85) pg。 结论 ①竹类植物叶片和笋均可作为基因组大小分析材料，细胞核染色时间对基因组大小测定结果存在一定影响，染色时间以3~5 min最佳。②热带木本竹种基因组大小明显小于温带木本竹种，温带木本竹种中，刚竹属竹种的基因组大小明显小于其他竹种。图1表5参29
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| 关 键 词： | 分子生物学 竹类植物 基因组大小 流式细胞仪 荧光峰值 |
| 收稿时间： | 2020-03-18 |
On the application of flow cytometry in the genomic size determination of bamboo plants |
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| CHEN Rongfen,HUANG Jianqin,CHEN Rong,XU Chuanmei.On the application of flow cytometry in the genomic size determination of bamboo plants[J].Journal of Zhejiang A&F University,2021,38(1):103-111. |
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| Authors: | CHEN Rongfen HUANG Jianqin CHEN Rong XU Chuanmei |
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| Affiliation: | State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China |
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| Abstract: | Objective]This research is aimed to study the effects of material tissue and the treatment method on the genome size of bamboo plants with the ultimate purpose to improve the genome size determination accuracy of bamboo plants.Method]With the leaves and shoots of different bamboo species selected as materials,and the rice used as the reference standard while the nuclear staining time set for 9 different gradients,that is 1,3,5,7,9,12,18,24 and 30 minutes,an investigation was carried out of the tissue sites and staining materials of different bamboo plants with the employment of flow cytometry.Result](1)With the same bamboo species,the leaves and shoots were similar in the fluorescence peak and genome size with the genome size difference range as narrow as 0.04~0.20 pg.(2)The 12 bamboo species are different in their nuclear staining time,with the fluorescence intensity of Sinobambusa tootsik,Sinobambusa tootisik f.albo-striata,Pseudosasa japonica var.tsutsumiana,Pseudosasa japonica f.akebono,Indocalamus decorus,Sasaella glabra f.albo-striata and Chimonobambusa mamorea f.variegata reaching the maximum within 1 minute,that of Bambusa multiplex and Phyllostachys sulphurea reaching the maximum within 3 minutes,that of Pseudosasa amabilis var.amabilis and Thyrsostachy ssiamensis reaching the maximum within 5 minutes,while that of Phyllostachys sulphurea(leaf)reaching the maximum within 7 minutes.(3)The fluorescence intensity of the 12 bamboos varies greatly from 1 to 30 minutes,all exceeding 5%except for the leaf of P.amabilis var.amabilis,T.ssiamensis,B.multiplex and the shoot of S.tootsik.In fact,the fluorescence intensity of P.japonica var.tsutsumiana and S.glabra f.albo-striata have reached 12.93%and 12.88%respectively.(4)As for the genome size of 12 bamboo species,2 tropical woody bamboo species of B.multiplex and T.ssiamensis changed from(2.64±0.54)pg to(2.69±1.01)pg,however,that of the 10 temperate woody bamboo species changed from(3.76±1.51)pg to(5.73±1.85)pg;of the 10 temperate bamboo species,the genome size of Phyllostachys changed from(3.76±1.51)pg to(3.91±0.95)pg,yet that of the other bamboo genus changed from(4.82±0.54)pg to(5.73±1.85)pg,which is obviously larger than Phyllostachys.Conclusion](1)Both leaves and shoots of bamboo can be used as experimental materials to determine their genome sizes by flow cytometry.The nuclear staining time has a certain effect on the determination of the bamboo genome size with 3 to 5 minutes as the optimal staining time.(2)The genome size of tropical woody bamboo species is obviously smaller than that of temperate woody bamboo species while among the temperate woody bamboo species,the genome size of Phyllostachys is obviously smaller than that of the other genus bamboo species. |
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| Keywords: | molecular biology bamboo plants genome size flow cytometry peak fluorescence |
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