Screening of Pear Genotypes with High-Efficiency Leaf Regeneration and Optimization of Leaf Regeneration System for‘Zaojinxiang’Pear

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

Acta Horticulturae Sinica ›› 2026, Vol. 53 ›› Issue (9): 2545-2559.doi: 10.16420/j.issn.0513-353x.2025-0836

• Genetic & Breeding · Germplasm Resources · Molecular Biology • Previous Articles Next Articles

Screening of Pear Genotypes with High-Efficiency Leaf Regeneration and Optimization of Leaf Regeneration System for‘Zaojinxiang’Pear

ZHANG Weifeng, WANG Fei, ZHANG Yanjie, QI Liyong, MA Li, LI Lixue, LUO Siming, YANG Zhen, JIANG Shuling^*(), OU Chunqing^*()

Key Laboratory of Biology and Genetic Improvement of Horticultural Crops， Germplasm Resources Utilization，Ministry of Agriculture and Rural Affairs，Research Institute of Pomology，Chinese Academy of Agricultural Sciences，Xingcheng， Liaoning 125100， China

Received:2025-11-07 Revised:2026-05-09 Online:2026-09-25 Published:2026-09-20
Contact: JIANG Shuling, OU Chunqing

Abstract

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

CLC Number:

S661.2

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.

Figures/Tables 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

Screening of Pear Genotypes with High-Efficiency Leaf Regeneration and Optimization of Leaf Regeneration System for‘Zaojinxiang’Pear

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品种或品系名称 Cultivar/Line name	存活外植体数/个 Number of surviving explants	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
B1-9	3	100.00	1.67
早金香 Zaojinxiang	18	72.22	1.92
早金香 Zaojinxiang^*	60	66.67	2.64
脆香 Cuixiang	7	71.43	2.20
山梨1 Pyrus ussuriensis 1	3	66.67	1.00
晚香 Wanxiang	18	55.56	1.30
SODAK	19	53.33	1.83
甘谷黑梨 Ganguheili	4	50.00	2.00
冬蜜 Dongmi	19	42.11	1.00
八月红 Bayuehong^*	60	41.70	1.49
甘长把 Ganchangba	18	33.33	1.00
洋梨2 Pyrus communis 2	3	33.33	1.00
金香水 Jinxiangshui	18	27.78	1.60
康弗伦斯 Conference	37	17.69	1.00
苹博香 Pingboxiang	9	11.11	1.00
2002-01-113	10	10.00	1.00

基本培养基 Basal medium	存活外植体数 Number of surviving explants	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
MS	35	85.98 ± 3.54 a	2.51 ± 0.35 ab
NN69	33	90.86 ± 0.84 a	2.64 ± 0.12 a
WPM	30	7.04 ± 6.12 b	1.00 ± 1.00 bc
1/2MS	30	13.33 ± 5.77 b	1.17 ± 0.29 c

TDZ/（mg · L^-1）	IAA/（mg · L^-1）	IBA/（mg · L^-1）	NAA/（mg · L^-1）	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
2.0	1.0			81.49 ± 16.96 ab	2.41 ± 0.94 a
2.0		0.3		62.93 ± 23.37 ab	1.71 ± 0.54 a
2.0			0.5	93.33 ± 5.77 a	2.50 ± 0.28 a
3.0	1.0			85.17 ± 9.49 ab	2.33 ± 0.35 a
3.0		0.3		53.02 ± 9.44 b	1.80 ± 0.35 a
3.0			0.5	68.85 ± 6.8 ab	1.55 ± 0.17 a

暗培养时间/d Dark culture time	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
7	50.00 ± 10.00 b	1.90 ± 0.66 a
14	93.30 ± 5.77 a	2.50 ± 0.28 a
21	76.67 ± 23.00 a	2.33 ± 0.29 a

叶片来源 Leaf source	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
2008年开始的组培苗Tissue cultured seedlings starting from 2008	69.76 ± 5.36 c	1.61 ± 0.35 b
2023年开始的组培苗Tissue cultured seedlings starting from 2023	79.24 ± 9.38 bc	2.39 ± 0.17 ab
2024年开始的组培苗Tissue cultured seedlings starting from 2024	87.22 ± 6.31 ac	2.52 ± 0.46 a
田间大树Big trees in the fields	96.97 ± 5.25 a	2.37 ± 0.15 ab
水培枝条Hydroponic shoots	93.64 ± 5.53 ab	2.51 ± 0.29 a

Kan/（mg · L^-1）	愈伤生长率/% Callus growth rate	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
0	100.00 ± 0.00 a	93.64 ± 5.53 a	2.51 ± 0.29 a
5	96.67 ± 5.77 a	66.67 ± 5.77 ab	3.16 ± 0.17 a
10	83.33 ± 5.77 a	50.00 ± 10.00 bc	2.46 ± 0.47 a
15	54.04 ± 11.17 b	32.63 ± 22.28 c	1.22 ± 0.19 b
20	33.53 ± 6.37 c	0 d	0 c

Cef /（mg · L^-1）	再生率/% Regeneration rate	平均再生芽数 Average number of regenerated buds
0	93.64 ± 5.53 a	2.51 ± 0.29 a
250	90.61 ± 9.11 a	2.89 ± 0.45 a
300	91.41 ± 8.35 a	2.71 ± 0.18 a
350	45.41 ± 7.96 b	1.30 ± 0.26 b
400	18.48 ± 8.64 c	1.00 ± 0.00 b

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