应用流式细胞技术鉴定桑树倍性的样品选择
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摘要
利用流式细胞仪对桑树不同叶位的叶片、同一叶位的不同部位、叶芽和种子胚根等进行倍性测定,筛选合适的桑树倍性鉴定材料。结果表明,未完全展开幼叶的检测效果优于第1叶位叶和第2叶位叶,第1叶位叶的检测效果优于第2叶位叶;同一叶位幼叶叶基部分的检测效果优于叶尾部分;叶芽的检测效果最好,而且制备样品的杂质较少,是用于流式细胞术研究的最佳材料。胚根也可以作为流式细胞仪检测的材料,这为更早检测桑树的倍性提供了候选材料。研究结果为选取不同时间段的不同材料进行桑树倍性检测提供了依据,也为其他植物倍性鉴定取材提供了参考。
In order to select suitable sample for ploidy identification of Morus alba L. more quickly and accurately, flow cytometry was adopted to identify ploidy of mulberry leaf from different positions, from different regions of the same leaf position, from leaf bud and seed radicle. Results showed that the test effect of the young leaf not fully expanded was better than that of the first leaf and the second leaf, the test effect of the first leaf was better than that of the second leaf, and the test effect of leaf base was better than that of leaf tail of the young leaves. Leaf bud with the best testing performance and the least impurities was the perfect material for ploidy identification of Morus alba L. by flow cytometry. In addition, the radicle can also be used for flow cytometry testing, providing potentials for early detection of mulberry ploidy. This experiment provides a basis for the sample selection of different materials in different time for mulberry ploidy detection, and also provides reference on sampling for ploidy identification of other plants.
In order to select suitable sample for ploidy identification of Morus alba L. more quickly and accurately, flow cytometry was adopted to identify ploidy of mulberry leaf from different positions, from different regions of the same leaf position, from leaf bud and seed radicle. Results showed that the test effect of the young leaf not fully expanded was better than that of the first leaf and the second leaf, the test effect of the first leaf was better than that of the second leaf, and the test effect of leaf base was better than that of leaf tail of the young leaves. Leaf bud with the best testing performance and the least impurities was the perfect material for ploidy identification of Morus alba L. by flow cytometry. In addition, the radicle can also be used for flow cytometry testing, providing potentials for early detection of mulberry ploidy. This experiment provides a basis for the sample selection of different materials in different time for mulberry ploidy detection, and also provides reference on sampling for ploidy identification of other plants.
引文
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[15] 杨静,宋勤霞,宁军权,等.利用流式细胞术鉴定桑树染色体倍性的方法[J].蚕业科学,2017,43(1):8-17
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[17] 翟晓巧,聂琳,张晓申.一种快速的木本植物倍性检测方法[J].陕西农业科学,2011,57(1):58-60
[2] 柯益富.桑树栽培及育种学[M].北京:农业出版社,1995:207
[3] 朱方容,雷扶生,胡乐山,等.四倍体桑研究和应用的概述[J].广西蚕业,1997(1):13-18
[4] 林强,朱方容,邱长玉,等.广西桑树四倍体资源的研究与应用[C]//中国蚕学会.中国蚕学会第六届青年学术研讨会论文集.杨凌:西北农林科技大学,2009:636-640
[5] 王颖,周攀,王玉霞,等.苹果属皱叶型植株实生后代倍性鉴定研究[J].北方园艺,2008(11):43-46
[6] EATON T D,CURLEY J,WILLIAMSON R C,et al.Determination of the level of variation in polyploidy among Kentucky bluegrass cultivars by means of flow cytometry[J].Crop Sci,2004,44(6):2168-2174
[7] GALBRAITH D W,HARKINS K R,MADDOX J M,et al.Rapid flow cytometric analysis of the cell cycle in intact plant tissues[J].Science,1983,220(4601):1049-1051
[8] 陈孟龙.草莓五倍体种间杂种染色体加倍及其十倍体株系生物学特性[D].南京:南京农业大学,2014
[9] 丁国昌,黄秀清,林思祖,等.流式细胞检测中杉木细胞核悬浮液的制备[J].西南林业大学学报,2013,33(1):97-100
[10] 胡磊.龙眼荔枝有性杂交、杂种流式细胞术鉴定及优株选育[D].广州:华南农业大学,2016
[11] 刘倩,魏臻武.流式细胞术分析苜蓿相对DNA含量的样品处理方法[J].草业科学,2014,31(9):1718-1723
[12] GALBRAITH D W.Flow cytometric analysis of plant genomes[J].Methods Cell Biol,1990,33(48):549-562
[13] 韩莎,李四军,许晓东,等.利用流式细胞仪快速鉴定桑树细胞的染色体倍数性[J].蚕业科学,2013,39(6):1042-1048
[14] 唐国华,贺修胜,龙治峰,等.流式细胞仪每秒进样细胞数对细胞周期分布的影响[J].中南医学科学杂志,2011,39(1):49-51
[15] 杨静,宋勤霞,宁军权,等.利用流式细胞术鉴定桑树染色体倍性的方法[J].蚕业科学,2017,43(1):8-17
[16] 曾民环,许德阳,薛景石,等.利用流式细胞仪研究拟南芥叶发育过程中细胞周期的调控[J].植物生理学报,2012,48(9):922-927
[17] 翟晓巧,聂琳,张晓申.一种快速的木本植物倍性检测方法[J].陕西农业科学,2011,57(1):58-60