基于SSR分子标记的山地刺槐克隆生长空间格局分析

doi:10.11707/j.1001-7488.LYKX20220483

Abstract

Abstract:

Objective: This study was carried out in black locust (Robinia pseudoacacia) forest in Huangtuliangzi, Pingquan City, Hebei Province, China, to verify the polymorphism of developed SSR primers of black locust, explore the distribution pattern of black locust sprouts, and provide a theoretical basis for artificial promotion of sprouting regeneration of black locust forests. Method: Three sample plots with obvious sprouting of black locust were selected in Huangtuliangzi forest farm of Pingquan City, Hebei Province. The genetic relationship of 125 black locust strains was identified by using 9 pairs of SSR primers. The traditional sample plot investigation method and the adjacent lattice method with variable scales were used to divide five sampling scales (5 m×5 m, 5 m×10 m, 10 m×10 m, 10 m×15 m, 20 m×30 m), calculate the deviation index (C) and its t-test, Cassie index (CA), average congestion (m*), negative binomial index (K), green index (GI) and agglomeration index (PAI), and then judge the pattern and scale. Result: The 9 pairs of primers had high polymorphism, and the average polymorphism information content was 0.76. In the three plots, a total of 73 genotypes, and 76 clonal ramets of black locust were identified. The distribution pattern of black locust clonal ramets from the cluster distribution tended to random distribution with the increase of sampling scale. The cluster distribution was the strongest when the sampling scale at 5 m×5 m. Conclusion: Based on the combination of SSR molecular markers and ecological investigation, this study explored a new method for studying the sprout distribution pattern, clarified the sprout distribution pattern of mountain black locust, and further understood the sprout regeneration theory of black locust. The distribution pattern of sprouts of black locust forest after felling is cluster distribution under no disturbance condition. The concentration and distribution of sprouting plants in a small area will result in a single genotype, which is not conducive to population diversity. It is suggested to make thinning of individuals that are too close to each other and affect normal growth and development, in order to ease competition and improve stand quality.

Key words: Robinia pseudoacacia, SSR, sprouting tillers, distribution pattern.

CLC Number:

Tianrun Cai,Jia Guo,Ziyi Wang,Yaxin Song,Shumin Zhang,Minsheng Yang,Jun Zhang. Spatial Pattern Analysis of Clonal Growth of Robinia pseudoacacia in Mountainous Areas Based on SSR Molecular Markers[J]. Scientia Silvae Sinicae, 2023, 59(6): 19-27.

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引物 Primer	正向引物序列 Forward primer sequence 5’→3’	反向引物序列 Reverse primer sequence 5’→3’	目的片段长度 Target fragment length/ bp	解链温度 Melting temperature(T_m)/ ℃
Rp-01	TGCAGAAAGAGAAAGCAGAGG	CCGAACCCTTTCTGGTTAGTC	140	53
Rp-04	CTCGTGATGATGGTGTTGATG	AATGGTCCAAACAACACGAAG	146	55
Rp-05	CCTTGCACATTTATCCCAGAA	CGACCTCGATCTTTTCTTGTG	158	55
Rp-06	TGGACAAAACATCATCGTGTG	CTCTCTTCTTTCTGCCCCTCA	147	55
Rp-12	AAGAGTCATCACGGAGACCAA	GGAGTCCAATTAAGTGCGAGA	150	55
Rp-15	TTAACTAATGCGGCGAGAAGA	GAGAGGAAGTGTGCGAAACAA	119	49
Rp-28	CTTGGTCTAGAAAGTCCTGCT	GGTCATCAAGGTTAGTTGGAT	151	55
Rp-32	CCACGTGGTTCTTCAAACATT	CAACAACAACCCACAAACACA	163	53
Rp-37	TGTCGTCATTTTATTTTACCC	CTCACCCTTTTTATTTCCATT	152	55

指标 Index	公式 Formula	判定标准 Judgment criteria
指标 Index	公式 Formula	集群分布 Cluster distribution	随机分布 Random distribution	均匀分布 Uniform distribution
偏离指数 Deviation index( C )	$C=S^2 / \bar{X} $ t检验 t-test： $t=(C-1) / \sqrt{[2 /(n-1)] }$ 自由度 Degree of freedom: n?1， t>t_0.05时达到显著水平 t>t_0.05reaching a significant level	C>1	C=1	C<1
负二项指数 Negative binomial distribution parameters( K )	$ K=\bar{X}^2 /\left(S^2-\bar{X}\right)$	K>0	K>8	K<0
Cassie 指标 Cassie index(CA)	CA=1/K	CA>0	CA=0	CA<0
平均拥挤度 Mean crowding( m*)	$m^*=C+\bar{X}-1 $	$m^{*} / \bar{X} > 1$	$m^* / \bar{X}=1 $	$m^* / \bar{X}<1 $
Green 指数 Green index(GI)	GI=(C?1)/(n?1)	GI>0	GI=0	GI<0
聚块性指标 Patchiness index(PAI)	PAI=1+1/K	PAI>1	PAI=1	PAI<1

引物 Primers	等位基因数目 No. of alleles	有效等位基因数目 No. of effective alleles	香农指数 Shannon's index	实际杂合度 Observed heterozygosity	预期杂合度 Expected heterozygosity	多态信息含量 Polymorphism information content	基因辨别能力 Probability of discrimination
RP-01	6	3.09	1.43	0.39	0.68	0.77	0.68
RP-04	7	6.26	1.89	0.49	0.84	0.90	0.84
RP-05	8	4.46	1.69	0.52	0.78	0.85	0.78
RP-06	9	4.71	1.72	0.38	0.79	0.88	0.79
RP-12	8	3.53	1.50	0.18	0.72	0.77	0.72
RP-15	4	2.15	1.02	0.30	0.53	0.60	0.53
RP-28	6	5.28	1.71	0.67	0.81	0.85	0.81
RP-32	6	2.26	1.00	0.07	0.56	0.56	0.56
RP-37	6	2.17	1.10	0.48	0.54	0.66	0.54
平均	5.89	4.19	1.52	0.33	0.74	0.76	0.74

样地 Plot	刺槐总数 Number of trees	有萌蘖刺槐数量 Number of cloned ramet genotypes	萌蘖刺槐总数 Total number of clone ramets	无萌蘖刺槐数量 Number of genotypes without clones	基因型总数 Number of genotypes	萌蘖刺槐占比 Proportion of cloned plants (%)
S1	33	6	18	15	21	54.6
S2	54	12	34	20	32	63.0
S3	38	6	24	14	20	63.2
总计 Total	125	24	76	49	73	60.8
均值 Average	41.67	8	25.33	16.33	26.33	60.2

格局参数 Pattern parameters	取样尺度 Sampling scale/
格局参数 Pattern parameters	5 m×5 m	5 m×10 m	10 m×10 m	10 m×15 m	20 m×30 m
单元数量 Unit number	83	41	19	13	3
偏离指数 Deviation index( C )	2.083	2.038	2.028	1.535	2.579
t	6.935*	4.643*	3.083*	1.311	1.579
t_0.05	1.989	2.026	2.110	2.201	4.303
负二项指数 Negative binomial distribution parameters( K )	0.845	1.786	3.892	10.926	16.044
Cassie指标 Cassie index(CA)	1.183	0.560	0.257	0.092	0.062
平均拥挤度 Mean crowding( m*)	1.999	2.892	5.028	6.381	26.912
Green指数 Green index	0.013	0.026	0.057	0.045	0.789
聚块性指标(GI) Patchiness index(PAI)	2.183	1.560	1.257	1.092	1.062
分布类型 Distribution type	集群分布 Cluster	集群分布 Cluster	集群分布 Cluster	随机分布Random	随机分布Random

Spatial Pattern Analysis of Clonal Growth of Robinia pseudoacacia in Mountainous Areas Based on SSR Molecular Markers

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[3]	Qingzhi Lin,Xiangyuan Zhu,Peili Mao,Lin Zhu,Longmei Guo,Zexiu Li,Banghua Cao,Yingdong Hao,Haitao Tan,Pizheng Hong,Xiaojun Lu. Effects of NaCl and PEG Stresses on Germination and Seedling Growth of Robinia pseudoacacia Seeds with Different Sizes [J]. Scientia Silvae Sinicae, 2022, 58(2): 100-112.
[4]	Lin Qi,Longmei Guo,Youde Liu,Banghua Cao,Peili Mao,Zexiu Li. Dynamic Responses of Non-Structural Carbohydrates in Robinia pseudoacacia Seedlings to NaCl Stress [J]. Scientia Silvae Sinicae, 2022, 58(1): 32-42.
[5]	Xin Peng,Hantang Wang,Chunhui Guo,Zhende Yang,Jing Zhou,Xue Wang,Zhirou Ding. EST-SSR Development and Cryptic Species Identification of the Invasive Gall-Causing Pest Leptocybe invasa (Hymenopetra: Eulophidae) [J]. Scientia Silvae Sinicae, 2021, 57(9): 140-151.
[6]	Jinfeng Cai,Xiaoming Yang,Wanwen Yu,Guibin Wang,Fuliang Cao. Development of SSR Molecular Markers Based on Transcriptome Sequencing of Melia azedarach [J]. Scientia Silvae Sinicae, 2021, 57(6): 85-92.
[7]	Chaoqun Du,Xiaomei Sun,Yunhui Xie,Yimei Hou. Genetic Diversity of Larix kaempferi Populations with Different Levels of Improvement in Northern Subtropical Region [J]. Scientia Silvae Sinicae, 2021, 57(5): 68-76.
[8]	Chao Liu,Lizhou Tang,Lihong Han. Characterization of the Chloroplast Genome of Lindera setchuenensis and Phylogenetics of the Genus Lindera [J]. Scientia Silvae Sinicae, 2021, 57(12): 167-174.
[9]	Yihong Meng,Gangbiao Xu,Mengzhu Lu,Xiaolong Jiang,Feilong Guo. Population Genetic Structure and Demographic History of Disanthus cercidifolius var. longipes [J]. Scientia Silvae Sinicae, 2020, 56(7): 55-62.
[10]	Maolin Yang,Xiaodong Ji,Heng Sun,Xu Cong,Guang Yang,Kai Hou,Yifan Ren. Comparation on Physical and Mechanical Properties of Branches, Stems and Roots of Robinia pseudoacacia at Different Ages [J]. Scientia Silvae Sinicae, 2020, 56(7): 115-122.
[11]	Xiaojun Zhou,Kai Zhang,Zhengfeng Peng,Shanshan Sun,Huiyuan Ya,Yanzhao Zhang,Yanwei Cheng. Comparative Analysis of Chloroplast Genome Characteristics between Paeonia jishanensis and Other Five Species of Paeonia [J]. Scientia Silvae Sinicae, 2020, 56(4): 82-88.
[12]	Jin Ling, Liu Mingguo, Dong Shengjun, Wu Yueliang, Zhang Xin. Genetic Diversity and Fingerprints of 97 Armeniaca sibirica Clones Based on SSR Markers [J]. Scientia Silvae Sinicae, 2018, 54(7): 51-61.
[13]	Bao Wenquan, Wuyun Tana, Du Hongyan, Li Tiezhu, Liu Huimin, Wang Lin, Bai Yu. Genetic Diversity and Population Structure of Amygdalus mira in the Tibet Plateau in China Based on SSR Markers [J]. Scientia Silvae Sinicae, 2018, 54(2): 30-41.
[14]	Li Hongguo, Xu Jihuang, Du Hongyan, Wuyun Tana, Liu Panfeng, Du Qingxin. Preliminary Construction of Core Collection of Eucommia ulmoides Based on Allele Number Maximization Strategy [J]. Scientia Silvae Sinicae, 2018, 54(2): 42-51.
[15]	Sun Huayu, Li Lichao, Zhao Hansheng, Yang Yihong, Wang Sining, Gao Zhimin. Molecular Characteristics and SSR Marker Development of Two NAC Genes from Daemonorops jenkinsiana [J]. Scientia Silvae Sinicae, 2017, 53(8): 132-140.