梨高效叶片再生基因型筛选与‘早金香’梨叶片再生体系优化

doi:10.16420/j.issn.0513-353x.2025-0836

摘要/Abstract

摘要：

以不同梨品种/品系不同时期叶片为外植体，经不同浓度次氯酸钠溶液消毒后进行叶片再生研究，以筛选高效叶片再生基因型，对筛选出来的再生率较高的‘早金香’的叶片再生条件进一步优化。结果表明，田间树和水培休眠枝条长出的梨幼叶均可用于梨叶片再生，消毒处理以2% NaClO处理7 min的效果最好，水培枝条长出的叶片存活率可达100%；供试82份梨品种/品系中仅15份获得了再生芽，且再生率多低于50%，其中‘B1-9’‘早金香’和‘脆香’3个品种的叶片再生率相对较高，分别为100%、72.22%和71.43%。对‘早金香’的叶片再生条件进一步优化，其中以田间幼叶在NN69 + 2.0 mg · L^-1 TDZ + 0.5 mg · L^-1 NAA + 蔗糖30 g · L^-1 + 琼脂8 g · L^-1的培养基上，暗培养14 d时的叶片再生效率最高，为96.97%。以卡那霉素和头孢霉素作为筛选和抑菌抗生素进行‘早金香’梨的遗传转化时，其最适浓度分别为20 mg · L^-1和250 ~ 300 mg · L^-1。

关键词: 梨, 叶片再生, 植物生长调节剂, 不定芽

Abstract:

To screen pear genotypes with high-efficiency leaf regeneration，the leaves of different pear varieties/lines at different stages were used as explants and disinfected by sodium hypochlorite solution with different concentrations，then the leaf regeneration was studied. ‘Zaojinxiang’was one of the screened genotypes with a relatively high leaf regeneration rate，its leaf regeneration conditions，concentrations of screening and antibacterial antibiotics were further optimized. The results indicated that tender pear leaves collected from the trees in orchard and the dormant shoots after water culture could be used for pear leaf regeneration. The disinfection treatment with 2% NaClO for 7 minutes achieved the best effect，and the explant survival rate of leaves from hydroponic shoots reached 100%. Only 15 of the 82 tested pear varieties/lines regenerated buds with the leaf regeneration rates were mostly below 50%. Among the regenerated varieties/lines，only three varieties/lines‘B1-9’‘Zaojinxiang’and‘Cuixiang’showed the relatively higher regeneration rates as 100%，72.22% and 71.43%，respectively. The leaf regeneration conditions of‘Zaojinxiang’were further optimized. The highest regeneration efficiency reached 96.97% when young leaves from the field were cultured in darkness for 14 days on a medium containing NN69 + 2.0 mg · L^-1 TDZ + 0.5 mg · L^-1 NAA + 30 g · L^-1 sucrose + 8 g · L^-1 agar. Using kanamycin and cefotaxime as screening and antibacterial antibiotics for the genetic transformation of‘Zaojinxiang’pear，the optimal concentrations were 20 mg · L^-1 and 250-300 mg · L^-1，respectively.

Key words: pear, leaf regeneration, plant growth regulator, adventitious bud

中图分类号:

S661.2

张薇丰, 王斐, 张艳杰, 齐力永, 马力, 李立学, 罗斯铭, 杨震, 姜淑苓, 欧春青. 梨高效叶片再生基因型筛选与‘早金香’梨叶片再生体系优化[J]. 园艺学报, 2026, 53(9): 2545-2559.

ZHANG Weifeng, WANG Fei, ZHANG Yanjie, QI Liyong, MA Li, LI Lixue, LUO Siming, YANG Zhen, JIANG Shuling, OU Chunqing. Screening of Pear Genotypes with High-Efficiency Leaf Regeneration and Optimization of Leaf Regeneration System for‘Zaojinxiang’Pear[J]. Acta Horticulturae Sinica, 2026, 53(9): 2545-2559.

图/表 12

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株系代号 Line code	亲本来源 Parental sources
S03-54、S03-62、S03-64、S03-75、S03-62、S03-105、S03-110、S03-115、S03-119、S03-124、S03-126、S03-128、S03-129、S05-101、S05-103、S05-110、S05-111和S05-114	锦丰 × 新水 Jinfeng × Shinsui
S06-94、S06-95、S06-96、S07-27、S07-44、S07-53、S07-54、S07-91、S07-71	火把 × 黄金 Huoba × Whangkeumbae
S08-9、S08-33、S08-43、S08-54、S08-56、S08-63、S08-72、S08-80、S09-40、S09-48、S09-46、S09-76、S09-82和S09-94	云南红梨 × 金花 Yunnanhongli × Jinhua
S10-16、S10-23、S10-24和S10-29	华酥 × 南果 Huasu × Nanguo
B1-9	红茄 × 黄冠 Red Clapp Favorite × Huangguan
11-7-3	中矮4号 × 南果 Zhongai 4 × Nanguo
08-7-120、08-7-128	中矮1号 × 日面红 Zhongai 1 × Flemish Beauty
08-6-112	锦丰 × 中梨一号 Jinfeng × Zhongli 1

株系代号 Line code	亲本来源 Parental sources
S03-54、S03-62、S03-64、S03-75、S03-62、S03-105、S03-110、S03-115、S03-119、S03-124、S03-126、S03-128、S03-129、S05-101、S05-103、S05-110、S05-111和S05-114	锦丰 × 新水 Jinfeng × Shinsui
S06-94、S06-95、S06-96、S07-27、S07-44、S07-53、S07-54、S07-91、S07-71	火把 × 黄金 Huoba × Whangkeumbae
S08-9、S08-33、S08-43、S08-54、S08-56、S08-63、S08-72、S08-80、S09-40、S09-48、S09-46、S09-76、S09-82和S09-94	云南红梨 × 金花 Yunnanhongli × Jinhua
S10-16、S10-23、S10-24和S10-29	华酥 × 南果 Huasu × Nanguo
B1-9	红茄 × 黄冠 Red Clapp Favorite × Huangguan
11-7-3	中矮4号 × 南果 Zhongai 4 × Nanguo
08-7-120、08-7-128	中矮1号 × 日面红 Zhongai 1 × Flemish Beauty
08-6-112	锦丰 × 中梨一号 Jinfeng × Zhongli 1

NaClO/%	处理时间/min Treatment time	无菌存活率/% Axenic survival rate	致死率/% Lethal rate	污染率/% Contamination rate
0.5	3	31.21 ± 4.66 df	6.06 ± 5.25 d	62.73 ± 7.77 a
	5	43.63 ± 3.15 cd	15.76 ± 5.85 cd	40.60 ± 4.58 bc
	8	6.67 ± 5.77 f	51.21 ± 11.87 ab	42.12 ± 7.06 b
1.0	5	52.07 ± 9.41 bd	11.36 ± 12.65 cd	36.56 ± 3.34 bc
	7	13.77 ± 7.33 ef	62.39 ± 15.60 a	23.84 ± 8.27 cf
	8	17.10 ± 7.03 ef	66.10 ± 9.23 a	16.80 ± 6.23 df
1.5	4	63.69 ± 7.88 abc	8.59 ± 8.35 d	27.73 ± 11.16 bf
	5	57.88 ± 7.06 abc	8.59 ± 8.35 d	33.53 ± 6.37 bcd
	6	59.93 ± 12.35 abc	10.72 ± 11.63 cd	29.35 ± 1.84 bf
	8	18.59 ± 10.59 ef	53.99 ± 12.4 ab	27.42 ± 2.50 bf
2.0	3	43.60 ± 6.86 cd	29.90 ± 8.70 bd	26.50 ± 3.95 bf
	4	73.74 ± 7.63 ab	2.78 ± 4.81 d	23.48 ± 4.73 cf
	5	28.52 ± 5.70 df	39.26 ± 5.59 abc	32.22 ± 1.92 bcde
	6	60.04 ± 8.64 abc	9.76 ± 9.17 cd	30.20 ± 3.04 bcde
	7	78.89 ± 11.70 a	5.56 ± 9.62 d	15.56 ± 5.09 ef
	8	32.99 ± 9.91 de	54.21 ± 9.61 ab	12.79 ± 4.77 f

NaClO/%	处理时间/min Treatment time	无菌存活率/% Axenic survival rate	致死率/% Lethal rate	污染率/% Contamination rate
0.5	3	31.21 ± 4.66 df	6.06 ± 5.25 d	62.73 ± 7.77 a
	5	43.63 ± 3.15 cd	15.76 ± 5.85 cd	40.60 ± 4.58 bc
	8	6.67 ± 5.77 f	51.21 ± 11.87 ab	42.12 ± 7.06 b
1.0	5	52.07 ± 9.41 bd	11.36 ± 12.65 cd	36.56 ± 3.34 bc
	7	13.77 ± 7.33 ef	62.39 ± 15.60 a	23.84 ± 8.27 cf
	8	17.10 ± 7.03 ef	66.10 ± 9.23 a	16.80 ± 6.23 df
1.5	4	63.69 ± 7.88 abc	8.59 ± 8.35 d	27.73 ± 11.16 bf
	5	57.88 ± 7.06 abc	8.59 ± 8.35 d	33.53 ± 6.37 bcd
	6	59.93 ± 12.35 abc	10.72 ± 11.63 cd	29.35 ± 1.84 bf
	8	18.59 ± 10.59 ef	53.99 ± 12.4 ab	27.42 ± 2.50 bf
2.0	3	43.60 ± 6.86 cd	29.90 ± 8.70 bd	26.50 ± 3.95 bf
	4	73.74 ± 7.63 ab	2.78 ± 4.81 d	23.48 ± 4.73 cf
	5	28.52 ± 5.70 df	39.26 ± 5.59 abc	32.22 ± 1.92 bcde
	6	60.04 ± 8.64 abc	9.76 ± 9.17 cd	30.20 ± 3.04 bcde
	7	78.89 ± 11.70 a	5.56 ± 9.62 d	15.56 ± 5.09 ef
	8	32.99 ± 9.91 de	54.21 ± 9.61 ab	12.79 ± 4.77 f

品种 Cultivar	NaClO/%	处理时间/min Treatment time	无菌存活率/% Axenic survival rate	污染率/% Contamination rate
早金香 Zaojinxiang	2	6	81.67 ± 11.69 b	18.33 ± 11.69 a
		7	100.00 ± 0.00 a	0 b
		8	100.00 ± 0.00 a	0 b
	3	6	93.33 ± 8.17 ab	6.67 ± 8.17 ab
		7	100.00 ± 0.00 a	0 b
		8	100.00 ± 0.00 a	0 b
八月红	2	6	86.67 ± 15.06 ab	13.33 ± 15.06 ab
Bayuehong		7	100.00 ± 0.00 a	0 b
		8	100.00 ± 0.00 a	0 b
	3	6	88.33 ± 13.29 ab	11.67 ± 13.29 ab
		7	100.00 ± 0.00 a	0 b
		8	100.00 ± 0.00 a	0 b