Bt转基因杨杂交子代苗期生长性状综合评价与筛选

doi:10.11707/j.1001-7488.LYKX20250637

Abstract

Abstract:

Objective: This study aims to combine the fast-growing traits of Populus ‘Nanlin 895’ with the high insect resistance of Bt-transgenic Populus nigra through hybridization, to analyze the effects of the Bt gene and sex on early growth, and to comprehensively evaluate the growth traits of the hybrid progeny for superior individual selection. Method: The ‘Nanlin 895’ poplar was used as the female parent and two Bt-transgenic Populus × euramericana clones (‘B3-44’ and ‘G-252’) were used as male parents, and two full-sib families were obtained through hybridization. Through seedling cultivation, the final effective sample size of 5 246 F₁ hybrid progeny were generated. During the dormancy period, plant height (PH), ground diameter (GD), and first-order branch number (FBN) were uniformly measured. PCR was employed for molecular identification of the Bt gene and progeny gender. The average membership function method and the Breckin multi-trait comprehensive evaluation method were used to comprehensively evaluate the growth traits. Result: 1) The three growth traits of the hybrid progeny exhibited abundant phenotypic variation, with coefficients of variation ranging from 20.3% to 48.3%. The FBN showed the highest level of variation. 2) The positive rate for the Bt gene was 35.3%. At seedling stage, the PH (270.66 cm) of Bt-positive progeny was significantly lower than that (274.23 cm) of Bt-negative progeny, but there were no significant differences in GD or FBN between Bt-positive progeny and Bt-negative progeny. 3) The gender ratio of the progeny was significantly skewed towards males, yet there were no significant differences in seedling growth traits between female and male progeny. 4) The top 100 superior individuals selected by the two comprehensive evaluation methods showed a high degree of overlap (96 individuals). Their mean PH and GD (351.58 cm, 45.48 mm) were significantly increased by 28.8% and 43.1%, respectively, compared to the population mean, and their FBN was more stable. Conclusion: The Bt gene has been successfully introduced into the poplar genome via hybridization, which may exert an inhibitory effect on seedling height growth to a certain extent. The gender of the progeny has no significant effect on early growth. Both the membership function and Breckin methods demonstrate good consistency and reliability for early selection in juvenile poplar. The superior individuals selected in this study provide a solid material foundation for breeding new catkin-free, fast-growing, and highly insect-resistant poplar varieties.

Key words: Populus spp., Bt gene, early sex determination, growth trait, comprehensive evaluation

CLC Number:

S722

Siyuan Li,Sheng Gao,Jianjun Hu,Jie Ling,Yingnan Chen. Comprehensive Evaluation and Selection of Growth Traits in Seedlings of Bt Transgenic Poplar Hybrid Progeny[J]. Scientia Silvae Sinicae, 2026, 62(6): 154-165.

Figures/Tables 12

Table 1

Table 2

Fig.S1

Fig.1

Fig.2

Fig.3

Table 3

Table 4

Fig.4

Table S1

Comprehensive evaluation and screening of superior individuals based on the membership function method"

子代编号Progeny no.	隶属函数值Membership value				排名Ranking
子代编号Progeny no.	株高PH	地径GD	一级分枝数FBN	均值Mean	排名Ranking
8-3-93	0.941	0.789	0.853	0.861	1
3-2-49	0.870	0.800	0.800	0.823	2
11-1-172	0.806	0.966	0.680	0.818	3
2-1-86	0.863	0.690	0.893	0.815	4
3-2-90	0.850	0.791	0.800	0.814	5
8-1-15	0.806	0.759	0.840	0.802	6
8-2-153	0.733	0.961	0.693	0.796	7
1-3-55	0.831	0.769	0.787	0.796	8
1-1-60	0.850	0.730	0.787	0.789	9
3-2-55	0.900	0.600	0.853	0.784	10
11-1-59	0.912	0.881	0.547	0.780	11
3-3-14	0.752	0.721	0.840	0.771	12
4-3-91	0.760	0.713	0.840	0.771	13
1-3-34	0.770	0.629	0.907	0.768	14
9-3-76	0.762	0.675	0.867	0.768	15
11-1-163	0.752	0.799	0.747	0.766	16
8-1-74	0.745	0.643	0.907	0.765	17
4-1-61	0.855	0.725	0.707	0.762	18
3-2-38	0.721	0.685	0.880	0.762	19
10-1-109	0.806	0.837	0.640	0.761	20
4-3-71	0.848	0.605	0.827	0.760	21
9-1-176	0.826	0.707	0.747	0.760	22
8-3-188	0.770	0.720	0.787	0.759	23
10-2-82	0.900	0.709	0.667	0.759	24
2-1-60	0.953	0.653	0.667	0.758	25
3-3-78	0.850	0.831	0.587	0.756	26
11-1-123	0.772	0.736	0.760	0.756	27
1-2-82	0.745	0.687	0.827	0.753	28
5-3-54	0.801	0.670	0.787	0.753	29
11-1-21	0.755	0.730	0.773	0.753	30
3-1-54	0.902	0.609	0.747	0.753	31
3-2-72	0.801	0.616	0.840	0.753	32
3-2-87	0.885	0.692	0.680	0.752	33
9-3-24	0.850	0.538	0.867	0.752	34
2-3-53	0.767	0.568	0.920	0.752	35
2-2-57	0.877	0.482	0.893	0.751	36
11-3-138	0.676	0.668	0.907	0.750	37
4-1-46	0.806	0.685	0.760	0.750	38
5-3-38	0.794	0.694	0.760	0.749	39
5-3-26	0.689	0.782	0.773	0.748	40
9-2-130	0.738	0.661	0.840	0.746	41
10-2-64	0.801	0.569	0.867	0.746	42
11-1-115	0.735	0.621	0.880	0.745	43
4-2-150	0.772	0.862	0.600	0.745	44
7-3-41	0.787	0.526	0.920	0.744	45
3-2-93	0.848	0.611	0.773	0.744	46
4-2-182	0.730	0.673	0.827	0.743	47
1-3-77	0.718	0.711	0.800	0.743	48
1-1-35	0.782	0.578	0.867	0.742	49
1-2-40	0.860	0.553	0.813	0.742	50
2-1-50	0.804	0.820	0.600	0.741	51
11-2-179	0.870	0.979	0.373	0.741	52
10-1-125	0.900	0.668	0.653	0.740	53
8-2-137	0.806	0.761	0.653	0.740	54
4-2-19	0.809	0.770	0.640	0.740	55
9-1-93	0.831	0.574	0.813	0.739	56
2-1-52	0.775	0.696	0.747	0.739	57
10-3-115	0.716	0.700	0.800	0.738	58
5-1-36	0.721	0.508	0.987	0.738	59
10-3-119	0.799	0.785	0.627	0.737	60
9-2-186	0.718	0.758	0.733	0.736	61
5-2-166	0.826	0.740	0.640	0.735	62
10-2-137	0.836	0.701	0.667	0.735	63
3-1-33	0.762	0.614	0.827	0.734	64
1-1-20	0.703	0.657	0.840	0.734	65
3-2-33	0.765	0.529	0.907	0.734	66
1-2-46	0.725	0.634	0.840	0.733	67
6-3-183	0.752	0.594	0.853	0.733	68
7-1-156	0.738	0.754	0.707	0.733	69
10-1-101	0.728	0.788	0.680	0.732	70
6-1-169	0.752	0.764	0.680	0.732	71
4-1-64	0.699	0.710	0.787	0.732	72
4-2-61	0.770	0.584	0.840	0.731	73
5-1-65	0.755	0.759	0.680	0.731	74
10-3-135	0.686	0.667	0.840	0.731	75
4-2-31	0.782	0.624	0.787	0.731	76
1-2-45	0.745	0.620	0.827	0.731	77
7-1-79	0.850	0.661	0.680	0.730	78
5-1-35	0.787	0.723	0.680	0.730	79
8-2-93	0.684	0.784	0.720	0.729	80
3-2-46	0.757	0.697	0.733	0.729	81
1-2-11	0.843	0.717	0.627	0.729	82
11-1-129	0.868	0.599	0.720	0.729	83
1-1-55	0.752	0.487	0.947	0.729	84
2-1-51	0.794	0.525	0.867	0.728	85
5-2-37	0.627	0.638	0.920	0.728	86
5-1-48	0.779	0.751	0.653	0.728	87
11-1-173	0.743	0.760	0.680	0.728	88
1-2-25	0.721	0.595	0.867	0.728	89
3-3-37	0.728	0.508	0.947	0.727	90
4-1-105	0.689	0.639	0.853	0.727	91
8-2-175	0.855	0.590	0.733	0.726	92
8-3-28	0.696	0.575	0.907	0.726	93
8-2-55	0.855	0.909	0.413	0.726	94
11-2-83	0.836	0.528	0.813	0.726	95
5-3-93	0.701	0.716	0.760	0.726	96
9-2-157	0.659	0.797	0.720	0.725	97
2-1-49	0.728	0.607	0.840	0.725	98
3-3-49	0.860	0.541	0.773	0.725	99
5-3-28	0.713	0.541	0.920	0.725	100

Table S1

Table S2

Superior individuals selected by Breckin multi-trait comprehensive evaluation method"

子代编号Progeny no.	各性状标准化得分Standardized score of each trait			综合评价值 Comprehensive evaluation value	排名Ranking
子代编号Progeny no.	株高PH	地径GD	一级分枝数FBN	综合评价值 Comprehensive evaluation value	排名Ranking
8-3-93	0.945	0.805	0.853	1.614	1
3-2-49	0.879	0.815	0.800	1.579	2
2-1-86	0.872	0.714	0.893	1.575	3
11-1-172	0.820	0.969	0.680	1.571	4
3-2-90	0.861	0.807	0.800	1.571	5
8-1-15	0.820	0.778	0.840	1.561	6
1-3-55	0.843	0.787	0.787	1.555	7
8-2-153	0.752	0.964	0.693	1.552	8
1-1-60	0.861	0.751	0.787	1.549	9
3-2-55	0.907	0.630	0.853	1.546	10
11-1-59	0.918	0.890	0.547	1.535	11
3-3-14	0.770	0.742	0.840	1.534	12
4-3-91	0.777	0.735	0.840	1.534	13
1-3-34	0.786	0.657	0.907	1.533	14
9-3-76	0.779	0.700	0.867	1.532	15
8-1-74	0.763	0.670	0.907	1.530	16
11-1-163	0.770	0.814	0.747	1.527	17
3-2-38	0.740	0.709	0.880	1.526	18
4-3-71	0.859	0.635	0.827	1.523	19
4-1-61	0.866	0.746	0.707	1.523	20
9-1-176	0.838	0.729	0.747	1.521	21
8-3-188	0.786	0.741	0.787	1.521	22
10-1-109	0.820	0.850	0.640	1.520	23
10-2-82	0.907	0.732	0.667	1.518	24
11-1-123	0.788	0.756	0.760	1.518	25
2-3-53	0.784	0.600	0.920	1.518	26
2-1-60	0.957	0.680	0.667	1.518	27
2-2-57	0.886	0.521	0.893	1.517	28
3-2-72	0.815	0.645	0.840	1.517	29
9-3-24	0.861	0.573	0.867	1.517	30
1-2-82	0.763	0.710	0.827	1.517	31
11-3-138	0.699	0.693	0.907	1.516	32
5-3-54	0.815	0.695	0.787	1.516	33
11-1-21	0.772	0.750	0.773	1.515	34
3-1-54	0.909	0.639	0.747	1.515	35
3-3-78	0.861	0.844	0.587	1.514	36
3-2-87	0.893	0.716	0.680	1.513	37
4-1-46	0.820	0.709	0.760	1.513	38
5-3-38	0.809	0.717	0.760	1.512	39
7-3-41	0.802	0.562	0.920	1.511	40
11-1-115	0.754	0.650	0.880	1.511	41
10-2-64	0.815	0.601	0.867	1.511	42
9-2-130	0.756	0.687	0.840	1.511	43
5-3-26	0.711	0.799	0.773	1.511	44
1-1-35	0.797	0.610	0.867	1.508	45
4-2-182	0.749	0.698	0.827	1.508	46
3-2-93	0.859	0.641	0.773	1.508	47
5-1-36	0.740	0.546	0.987	1.507	48
1-3-77	0.738	0.733	0.800	1.507	49
1-2-40	0.870	0.587	0.813	1.507	50
9-1-93	0.843	0.607	0.813	1.504	51
4-2-150	0.788	0.872	0.600	1.503	52
10-3-115	0.736	0.723	0.800	1.503	53
2-1-52	0.790	0.720	0.747	1.502	54
10-1-125	0.907	0.693	0.653	1.501	55
3-2-33	0.781	0.565	0.907	1.501	56
8-2-137	0.820	0.779	0.653	1.501	57
2-1-50	0.818	0.834	0.600	1.501	58
4-2-19	0.822	0.788	0.640	1.500	59
3-1-33	0.779	0.644	0.827	1.500	60
6-3-183	0.770	0.625	0.853	1.499	61
1-1-20	0.724	0.684	0.840	1.499	62
9-2-186	0.738	0.776	0.733	1.499	63
1-2-46	0.745	0.662	0.840	1.499	64
1-1-55	0.770	0.526	0.947	1.497	65
4-2-61	0.786	0.616	0.840	1.497	66
10-3-119	0.813	0.801	0.627	1.497	67
10-3-135	0.708	0.692	0.840	1.497	68
3-3-37	0.747	0.545	0.947	1.496	69
1-2-45	0.763	0.649	0.827	1.496	70
5-2-37	0.654	0.665	0.920	1.496	71
4-1-64	0.720	0.732	0.787	1.496	72
5-2-166	0.838	0.760	0.640	1.496	73
10-2-137	0.847	0.724	0.667	1.496	74
4-2-31	0.797	0.653	0.787	1.496	75
2-1-51	0.809	0.561	0.867	1.495	76
7-1-156	0.756	0.773	0.707	1.495	77
11-2-179	0.879	0.981	0.373	1.494	78
1-2-25	0.740	0.626	0.867	1.494	79
8-3-28	0.718	0.608	0.907	1.494	80
10-1-101	0.747	0.805	0.680	1.494	81
6-1-169	0.770	0.782	0.680	1.494	82
4-1-105	0.711	0.667	0.853	1.494	83
5-3-28	0.733	0.576	0.920	1.493	84
5-1-65	0.772	0.777	0.680	1.493	85
3-2-46	0.774	0.720	0.733	1.493	86
7-1-79	0.861	0.687	0.680	1.493	87
8-2-93	0.706	0.801	0.720	1.492	88
11-1-129	0.877	0.630	0.720	1.492	89
3-3-22	0.661	0.685	0.880	1.492	90
5-1-35	0.802	0.744	0.680	1.492	91
11-2-83	0.847	0.564	0.813	1.492	92
2-1-49	0.747	0.637	0.840	1.491	93
4-1-24	0.765	0.603	0.853	1.490	94
8-2-65	0.843	0.511	0.867	1.490	95
8-2-175	0.866	0.621	0.733	1.490	96
1-3-49	0.786	0.568	0.867	1.490	97
5-3-93	0.722	0.738	0.760	1.490	98
1-2-11	0.854	0.739	0.627	1.490	99
3-3-49	0.870	0.576	0.773	1.490	100

Table S2

Table S3

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引物名称 Primer name	引物序列 Primer sequence （5’→3’）	产物长度 Product length/bp	反应条件 Reaction conditions
Cry1Ac_F	GAATTCGCTAGGAACCAAGCCATT	633	94 ℃ 5 min 94 ℃ 30 s 57 ℃ 30 s 72 ℃ 30 s 72 ℃ 7 min
Cry1Ac_R	AAGTAT ATCCATCAAATGTGGACT	633	94 ℃ 5 min 94 ℃ 30 s 57 ℃ 30 s 72 ℃ 30 s 72 ℃ 7 min
MSL_F	AGCAGTATTTATAGGCTAGAGG	399	94 ℃ 5 min 94 ℃ 30 s 57 ℃ 30 s 72 ℃ 30 s 72 ℃ 7 min
MSL_R	CATTGTAATAACATTTCTTGCTC	399	94 ℃ 5 min 94 ℃ 30 s 57 ℃ 30 s 72 ℃ 30 s 72 ℃ 7 min

生长性状 Growth trait	平均值 Mean	最大值 Max.	最小值 Min.	标准差SD	变异系数 CV (%)	正态性检验 Norm test (W)
株高PH/cm	272.97	439	31	55.42	20.3	0.97
地径GD/mm	31.78	64.93	4.95	9.02	28.4	0.99
一级分枝数FBN	23.18	75	0	11.2	48.3	0.99

生长性状 Growth trait	Bt阳性子代 Bt positive progeny (n=1 850)			Bt阴性子代 Bt negative progeny (n=3 396)			t 检验 t-test
生长性状 Growth trait	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV (%)	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV (%)	t	P
株高PH/cm	270.66±54.77	31~439	20.2	274.23±55.73	60~420	20.3	?2.23	0.03
地径GD/mm	31.63±9.02	5.98~62.59	28.5	31.86±9.01	4.95~64.93	28.3	?0.90	0.37
一级分枝数FBN	23.27±11.38	0~75	48.9	23.12±11.10	0~74	48.0	0.45	0.65

生长性状 Growth trait	雌株子代 Female progeny (n=2 390)			雄株子代 Male progeny (n=2 856)			t 检验 t-test
生长性状 Growth trait	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV (%)	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV (%)	t	P
株高PH/cm	272.02±56.43	31~439	20.7	273.77±54.56	60~415	19.9	1.13	0.26
地径GD/mm	31.53±9.09	4.95~63.12	28.9	31.99±8.95	5.26~64.93	28.0	1.84	0.07
一级分枝数FBN	23.38±11.23	0~75	48.0	23.01±11.17	0~74	48.5	?1.19	0.24

生长性状 Growth trait	隶属函数法 Membership function method			布雷金多性状综合评定法 Breckin multi-trait comprehensive evaluation method
生长性状 Growth trait	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV	均值±标准差 Mean ± SD	变异范围 Range	变异系数 CV
株高 PH （cm）	351.80 ± 27.17	287-420	7.72%	351.58 ± 27.29	287-420	7.76%
地径 GD （mm）	46.07 ± 6.33	33.86-63.68	13.74%	45.48 ± 6.34	33.18-63.68	13.93%
一级分枝数 FBN （no./plant）	16.93± 8.22	1-47	48.54%	16.18± 7.75	1-47	47.88%

Comprehensive Evaluation and Selection of Growth Traits in Seedlings of Bt Transgenic Poplar Hybrid Progeny

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