Establishment and Application of Ploidy Method for the Identification of Psidium guajava by Flow Cytometry

doi:10.13560/j.cnki.biotech.bull.1985.2023-0844

Abstract

Abstract:

【Objective】 The flow cytometry was used to identify the ploidy of Psidium guajava, which may lay the foundation for ploidy breeding and hybrid breeding of P. guajava. 【Method】 Using P. guajava as the material, the effects of sample parts, centrifugation treatment, and storage methods on ploidy detection results were compared. 【Result】 P. guajava petals had the highest number of collected nuclear DNA, with a CV value of 1.96% and the best peak shape. When preparing nuclear suspension, the best detection effect was achieved by directly staining and analyzing without centrifugation after filtration. The nuclear DNA content obtained from fresh P. guajava petals was significantly higher than that of refrigerated and other frozen treatment groups, with fewer background fragments and a clear main peak. The -80℃ freezing treatment caused the least damage to the samples, and the detection effect was second only to that of fresh P. guajava petals. The established flow cytometry method for ploidy analysis of P. guajava involved taking 0.50-1.00 cm² of pomegranate petals, mixing them with 1 mL of mGb lysis buffer, filtering, and adding 30 μL of PI staining for 1 min before analysis. 【Conclusion】 Using this established flow cytometry method, ploidy identification was performed on 33 P. guajava germplasm resources, resulting in the detection of 32 diploids and 1 hexaploid. This method is simple, efficient, and accurate, and may provide an effective approach for ploidy identification of P. guajava germplasm resources.

Key words: Psidium guajava, petal, ploidy identification, flow cytometry

SHAO Xue-hua, LI Gui-lan, XIAO Wei-qiang, LAI Duo, ZHUANG Qing-li, QIN Jian. Establishment and Application of Ploidy Method for the Identification of Psidium guajava by Flow Cytometry[J]. Biotechnology Bulletin, 2024, 40(2): 48-54.

Figures/Tables 5

References 23

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品种编号Sample No.	名称Name	倍性系数Coefficient of ploidy	倍性估算值Ploidy estimated value	倍性Ploidy
1	紫果Ziguo	0.49	1.96	2n
2	泰国2号Thailand 2	0.45	1.80	2n
3	泰国3号Thailand 3	0.48	1.92	2n
4	软香Ruanxiang	0.43	1.72	2n
5	木瓜Mugua	0.53	2.12	2n
6	红肉实生Hongrou shisheng	0.59	2.36	2n
7	红肉翁拔Hongrouwengba	0.51	2.04	2n
8	帝王Diwang	0.48	1.92	2n
9	草莓Strawberry	0.52	2.01	6n
10	西瓜红Xiguahong	0.44	1.76	2n
11	东莞红肉Dongguanhongrou	0.45	1.80	2n
12	佳硕Jiashuo	0.46	1.84	2n
13	水蜜Shuimi	0.49	1.96	2n
14	细叶小果Xiyexiaoguo	0.44	1.76	2n
15	细叶大果Xiyedaguo	0.43	1.72	2n
16	金斗香Jin douxiang	0.45	1.80	2n
17	白肉翁拔Bairouwengba	0.52	2.08	2n
18	翡翠Feicui	0.51	2.04	2n
19	青皮红肉Qingpihongrou	0.53	2.12	2n
20	金石宫Jinshigong	0.47	1.88	2n
21	珍珠Zhenzhu	0.50	2.00	2n
22	红心软果Hongxinruanguo	0.42	1.68	2n
23	粉红蜜Fenhongmi	0.51	2.04	2n
24	卵红Luanhong	0.50	2.00	2n
25	宫粉红Gongfenhong	0.58	2.32	2n
26	黄皮红肉Huangpihongrou	0.53	2.12	2n
27	B1	0.51	2.04	2n
28	B2	0.56	2.24	2n
29	B3	0.51	2.04	2n
30	B4	0.54	2.16	2n
31	B5	0.53	2.12	2n
32	B6	0.53	2.12	2n
33	穗红1号Suihong 1	0.49	1.96	2n

品种编号Sample No.	名称Name	倍性系数Coefficient of ploidy	倍性估算值Ploidy estimated value	倍性Ploidy
1	紫果Ziguo	0.49	1.96	2n
2	泰国2号Thailand 2	0.45	1.80	2n
3	泰国3号Thailand 3	0.48	1.92	2n
4	软香Ruanxiang	0.43	1.72	2n
5	木瓜Mugua	0.53	2.12	2n
6	红肉实生Hongrou shisheng	0.59	2.36	2n
7	红肉翁拔Hongrouwengba	0.51	2.04	2n
8	帝王Diwang	0.48	1.92	2n
9	草莓Strawberry	0.52	2.01	6n
10	西瓜红Xiguahong	0.44	1.76	2n
11	东莞红肉Dongguanhongrou	0.45	1.80	2n
12	佳硕Jiashuo	0.46	1.84	2n
13	水蜜Shuimi	0.49	1.96	2n
14	细叶小果Xiyexiaoguo	0.44	1.76	2n
15	细叶大果Xiyedaguo	0.43	1.72	2n
16	金斗香Jin douxiang	0.45	1.80	2n
17	白肉翁拔Bairouwengba	0.52	2.08	2n
18	翡翠Feicui	0.51	2.04	2n
19	青皮红肉Qingpihongrou	0.53	2.12	2n
20	金石宫Jinshigong	0.47	1.88	2n
21	珍珠Zhenzhu	0.50	2.00	2n
22	红心软果Hongxinruanguo	0.42	1.68	2n
23	粉红蜜Fenhongmi	0.51	2.04	2n
24	卵红Luanhong	0.50	2.00	2n
25	宫粉红Gongfenhong	0.58	2.32	2n
26	黄皮红肉Huangpihongrou	0.53	2.12	2n
27	B1	0.51	2.04	2n
28	B2	0.56	2.24	2n
29	B3	0.51	2.04	2n
30	B4	0.54	2.16	2n
31	B5	0.53	2.12	2n
32	B6	0.53	2.12	2n
33	穗红1号Suihong 1	0.49	1.96	2n