摘要: 目的‘京�?9’是一种优质的枣鲜食新品种，目前尚无有效的离体再生体系，影响了自根苗的培育和苗木纯度。为了解决枣遗传转化效率低等问题，本研究探究了枣叶片再生的不同影响因素，以期建立高效的叶片再生体系、�/sec> 方法以‘京�?9’组培苗叶片为试材，研究了不同接种方式、基本培养基种类、植物生长调节剂种类和浓度配比对离体叶片直接诱导不定芽再生的影响、�/sec> 结果正交试验结果表明：叶片正放的接种方式再生率远高于叶片反放，是‘京�?9’叶片再生不定芽的最佳放置方式；基本培养基对‘京�?9’叶片再生的影响要大于植物生长调节剂，且最优培养基�?/2 MS，最佳植物生长调节剂为细胞分裂素6-BA，本研究确立的‘京�?9’组培苗叶片再生的最优体系为1/2 MS + 1 mg/L噻苯� + 0.5 mg/L 6-BA + 0.2 mg/L NAA + 30 g/L麦芽� + 6 g/L琼脂，再生率可达79.17%，平均每叶片再生不定芽能达到4.67个。进而初探并建立了‘京�?9’组培苗的最佳扩繁体系为：MS + 20 g/L麦芽� + 6 g/L琼脂 + 3 mg/L 6-BA + 0.1 mg/L NAA，分枝率高达88%，平均分枝数�?.76枝；以及最适生根体系为�?/2MS + 20 g/L蔗糖 + 8 g/L琼脂 + 1.5 mg/L IBA + 0.3 mg/L NAA，生根率�?0.91%，平均根长达4.08 cm、�/sec> 结论‘京�?9’叶片再生效率受外界影响因素很大，尤其是基本培养基种类和接种方式，且每个因素都不是独立起作用的。本研究为之后以‘京�?9’叶片为转基因材料进行枣相关基因功能验证建立了完整的培育体系，同时为加快枣的相关研究进程奠定基础、�/sec>

关键诌�
• ‘京�?9’叶牆�/a> /
• 直接再生/
• 不定芼�/a> /
• 最优培养基

Abstract: Objective‘Jingzao 39� is a new high-quality jujube variety. There is no effective regeneration system in vitro at present, which affects the cultivation of self-rooted seedlings and the purity of seedlings. In order to solve the problems of low genetic transformation efficiency of jujube, different influencing factors of jujube leaf regeneration were explored in this study, in order to establish an efficient leaf regeneration system. MethodUsing the leaves of tissue cultured seedlings of ‘Jingzao 39� as experimental materials, the effects of different inoculation methods, types of basic culture medium, types and concentration ratio of plant growth regulators on the direct induction of adventitious bud regeneration from detached leaves were studied. ResultThe results showed that the regeneration rate of the inoculation method with the leaves being placed upward was much higher than that of back up, which was the best placement method for the regeneration of adventitious buds from leaves of ‘Jingzao 39�? The effect of basic culture medium on leaf regeneration of ‘Jingzao 39� was greater than that of plant growth regulator, and the optimal basic medium and plant growth regulator were 1/2 MS and cytokinin 6-BA, respectively. The optimal regeneration system for leaves of ‘Jingzao 39� tissue cultured seedlings established in this study was 1/2 MS medium + 1.0 mg/L thiabenduron + 0.5 mg/L 6-BA + 0.2 mg/L NAA + 30 g/L maltose + 6 g/L agar, an average of 4.67 adventitious buds could be regenerated for each leaf. Then, the optimal propagation system of ‘Jingzao 39� tissue culture seedlings was initially explored and established: MS + 20 g/L maltose + 6 g/L agar + 3.00 mg/L 6-BA + 0.10 mg/L NAA, with the branching rate up to 88%, and the average branch number of 2.76; the most suitable rooting medium was: 1/2MS + 20 g/L sucrose + 8 g/L agar + 1.50 mg/L IBA + 0.30 mg/L NAA, with rooting rate of 90.91%, and the average root length up to 4.08 cm. ConclusionThe leaf regeneration efficiency of ‘Jingzao 39� is greatly influenced by external factors, especially the basic medium type and inoculation method, and each factor does not play an independent role. This study establishes a complete cultivation system for the functional verification of interested gene with the leaves of ‘Jingzao 39� as transgenic materials, and lays a foundation for accelerating the jujube research process.

Key words:
• ‘Jingzao 39� leaf/
• direct regeneration/
• adventitious bud/
• optimal medium

�?nbsp; 1J39离体叶片�?种培养基中不同接种方式直接再生不定芽示意国�/p>

A ~ C. 离体叶片分别�?�?1�?2号培养基中的初始诱导状；D ~ E. 叶片分别�?�?1�?2号培养基中暗培养14 d后的状态；G ~ I. 叶片分别�?�?1�?2号培养基以及2种接种方式下分化出不定芽，其中红箭头所指为叶片正放，其他为反放。A−C, the initial induction state of detached leaves in medium 2, 11, and 12; D−E, the state of leaves after dark culture in medium 2, 11, and 12 for 14 days; G−I, the leaves are differentiated into adventitious buds on medium 2, 11, and 12 under the two inoculation methods: the red arrow points to the leaf being placed upright and the other is reversed.

Figure 1.Schematic diagram of adventitious buds directly regenerated from J39 detached leaves in three media with different inoculation methods

下载: 全尺寸图牆�/a> 幻灯牆�/a>

�?nbsp; 2无菌苗在3种糖源的扩繁体系中生长情冴�/p>

A ~ B. 糖源�?0 g/L蔗糖；C ~ D. 糖源�?5 g/L麦芽糖和7.50 g/L蔗糖；E ~ F. 糖源�?0 g/L麦芽糖。A−B, the source of sugar is 20 g/L sucrose; C−D, the sugar source is 15 g/L maltose and 7.50 g/L sucrose; E−F, the source of sugar is 20 g/L maltose.

Figure 2.Growth condition of sterile seedlings in the propagation system of three glycogens

下载: 全尺寸图牆�/a> 幻灯牆�/a>

�?nbsp; 3无菌苗的生根情况

A. 无菌苗接种初期的基部形态；B. 粗壮根；C. 纤细根；D. 根上长出须根。A, the basal morphology at the initial stage of inoculation; B, thick roots; C, slender roots; D, fibrous roots growing on the roots.

Figure 3.Rooting situation of aseptic seedlings

下载: 全尺寸图牆�/a> 幻灯牆�/a>

�?nbsp; 1离体叶片不定芽诱导再生正交试骋�/p>

Table 1.Orthogonal test of induction of adventitious bud regeneration from detached leaves

水平 Level	培养基种籺�br/>Type of medium ＇�i>A（�/td>	TDZ＇�i>B�?（mg·L�?（�/td>	NAA＇�i>C�?（mg·L�?（�/td>	6-BA＇�i>D�?（mg·L�?（�/td>	IBA＇�i>E�?（mg·L�?（�/td>
1	1/2 MS	0.00	0.00	0.00	0.00
2	MS	0.50	0.20	0.50	0.20
3	WPM	1.00	0.40	1.00	0.40

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�?nbsp; 2离体叶片不定芽诱导再生正交试验因素水干�/p>

Table 2.Orthogonal test factor level of adventitious bud induction and regeneration from detached leaves

水平 Level	糖源 Glycogen ＇�i>a（�/td>	6-BA＇�i>b�?（mg·L�?（�/td>	IBA＇�i>c�?（mg·L�?（�/td>	NAA＇�i>d�?（mg·L�?（�/td>
1	蔗糖20.00 g/L Sucrose 20.00 g/L	1.00	0.00	0.00
2	麦芽�?5.00 g/L + 蔗糖7.50 g/L Maltose 15.00 g/L + sucrose 7.50 g/L	2.00	0.40	0.10
3	麦芽�?0.00 g/L Maltose 20.00 g/L	3.00	0.80	0.20

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�?nbsp; 3不同接种方式对J39叶片再生的影哌�/p>

Table 3.Effects of different inoculation methods on leaf regeneration of ‘Jingzao 39‘�/p>

试验叶�br/>Test No.	接种方式 Vaccination method	接种叶片�?br/>Number of inoculated leaf	平均芽数 Average number of bud
2	叶正面朝上（正） Leaf face up (positive)	12	2.31
	叶背面朝上（反） Leaf dorsal facing up (reverse)	12	0.52
11	叶正面朝上（正） Leaf face up (positive)	12	1.93
	叶背面朝上（反） Leaf dorsal facing up (reverse)	12	0.67
12	叶正面朝上（正） Leaf face up (positive（�/td>	12	4.79
	叶背面朝上（反） Leaf dorsal facing up (reverse)	12	0.71

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�?nbsp; 4离体叶片不定芽诱导再生正交试验结枛�/p>

Table 4.Orthogonal test results of adventitious bud induced regeneration of detached leaves

试验叶�br/>Test No.	A	B/(mg·L�?)	C/(mg·L�?)	D/(mg·L�?)	E/(mg·L�?)	平均芽数 Average number of bud
1	1/2 MS	0.00	0.00	0.00	0.00	0.00
2	1/2 MS	0.50	0.20	0.50	0.20	2.24
3	1/2 MS	1.00	0.40	1.00	0.40	0.33
4	MS	0.00	0.00	0.50	0.20	0.00
5	MS	0.50	0.20	1.00	0.40	0.00
6	MS	1.00	0.40	0.00	0.00	0.00
7	WPM	0.00	0.20	0.00	0.40	0.00
8	WPM	0.50	0.40	0.50	0.00	0.00
9	WPM	1.00	0.00	1.00	0.20	0.23
10	1/2 MS	0.00	0.40	1.00	0.20	0.18
11	1/2 MS	0.50	0.00	0.00	0.40	2.22
12	1/2 MS	1.00	0.20	0.50	0.00	4.67
13	MS	0.00	0.20	1.00	0.00	0.00
14	MS	0.50	0.40	0.00	0.20	0.46
15	MS	1.00	0.00	0.50	0.40	0.08
16	WPM	0.00	0.40	0.50	0.40	0.00
17	WPM	0.50	0.00	1.00	0.00	0.04
18	WPM	1.00	0.20	0.00	0.20	0.00
K1	9.64	0.18	2.58	2.68	4.71
K2	0.54	4.97	6.91	6.99	3.11
K3	0.27	5.31	0.97	0.78	2.64
x1	1.60	0.03	0.43	0.45	0.78
x2	0.09	0.83	1.15	1.17	0.52
x3	0.05	0.89	0.16	0.13	0.44
R	1.56	0.86	0.99	1.03	0.34
注：Ki= Σ（该因素�?i>i水平下的平均芽数），xi=Ki/n＇�i>n= 该水平的重复数）＋�i>R为极差。下同。Note:Ki= Σ (the average number of buds of the factor at theilevel),xi=Ki/n(n= the number of replicates at this level),Rmeans range. The same below.

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�?nbsp; 5不同因素对J39不定芽再生数影响的极差分枏�/p>

Table 5.Range analysis of effects of different factors on the regeneration number of J39 adventitious buds

因子 Factor	水平1 Level 1	水平2 Level 2	水平3 Level 3	R
A	1.61	0.09	0.05	1.56
B	0.03	0.83	0.89	0.86
C	0.43	1.15	0.16	0.99
D	0.45	1.17	0.13	1.03
E	0.78	0.52	0.44	0.34

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�?nbsp; 6不同因素对J39不定芽再生数影响的方差分枏�/p>

Table 6.Variance analysis of effects of different factors on the regeneration number of J39 adventitious buds

因子 Factor	SS	df	MS	F	P
A	9.48	2	4.74	4.77*	0.05
B	2.75	2	1.37	1.38	0.31
C	3.14	2	1.57	1.58	0.27
D	3.37	2	1.69	1.70	0.25
E	0.39	2	0.20	0.20	0.83
误差 Error	6.95	7	1.00
总计 Total	26.08	17
注：F� >F0.05表示显著差异，数值右上角标注�?”。Notes:Fvalue >F0.05means significant difference, with a �?� in the upper right corner of the value.

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�?nbsp; 73种基本培养基对J39不定芽再生数影响的LSD多重比较

Table 7.Multiple comparisons of LSD of the effects of 3 basic media on the regeneration number of J39 adventitious buds

基本培养培�br/>Basic medium	基本培养培�br/>Basic medium	平均值差� Mean difference	标准�?br/>Standard error	显著�?br/>Significance	95% 置信区间 95% confidence interval
					下限 Lower limit	上限 Upper limit
1/2 MS	MS	1.52*	0.61	0.03	0.22	2.81
	WPM	1.56*	0.61	0.02	0.27	2.86
MS	1/2 MS	�?.52*	0.61	0.03	�?.81	�?.22
	WPM	0.05	0.61	0.94	�?.25	1.34
WPM	1/2 MS	�?.56*	0.61	0.02	�?.86	�?.27
	MS	�?.05	0.61	0.94	�?.34	1.25
注：*显著性水平为0.05。Note:*means the significance level of 0.05.

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�?nbsp; 8J39幼苗扩繁正交试验结果

Table 8.Orthogonal test results of J39 seedling proliferation

试验叶�br/>Test No.	a	b/（mg·L�?（�/td>	c/（mg·L�?（�/td>	d/（mg·L�?（�/td>	分枝玆�br/>Branching rate/%
1	蔗糖20.00 g/L Sucrose 20.00 g/L	1.00	0.00	0.00	47.37
2	蔗糖20.00 g/L Sucrose 20.00 g/L	2.00	0.40	0.10	85.00
3	蔗糖20.00 g/L Sucrose 20.00 g/L	3.00	0.80	0.20	68.42
4	麦芽�?5.00 g/L + 蔗糖7.50 g/L Maltose 15.00 g/L + sucrose 7.50 g/L	1.00	0.40	0.20	40.00
5	麦芽�?5.00 g/L + 蔗糖7.50 g/L Maltose 15.00 g/L + sucrose 7.50 g/L	2.00	0.80	0.00	56.52
6	麦芽�?5.00 g/L + 蔗糖7.50 g/L Maltose 15.00 g/L + sucrose 7.50 g/L	3.00	0.00	0.10	80.00
7	麦芽�?0.00 g/L Maltose 20.00 g/L	1.00	0.80	0.10	56.52
8	麦芽�?0.00 g/L Maltose 20.00 g/L	2.00	0.00	0.20	78.26
9	麦芽�?0.00 g/L Maltose 20.00 g/L	3.00	0.40	0.00	71.43
K1	200.79	143.89	205.63	175.32
K2	176.52	219.78	196.43	221.52
K3	206.21	219.85	181.46	186.68
x1	66.93	47.96	68.54	58.44
x2	58.84	73.26	65.48	73.84
x3	68.74	73.28	60.49	62.23
R	9.90	25.32	8.05	15.40

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�?nbsp; 9无菌苗在不同培养基中不定根的诱导情况

Table 9.Adventitious root induction of aseptic seedlings on different media

培养基号 Medium No.	蔗糖 Sucrose/（g·L�?（�/td>	IBA/（mg·L�?（�/td>	NAA/（mg·L�?（�/td>	生根玆�br/>Rooting rate/%	平均根长 Average root length/cm
1	10	0.50	0.00	62.50cd	3.68def
2	20	0.50	0.00	66.67bcd	5.03cd
3	30	0.50	0.00	60.00cd	2.73f
4	10	0.50	0.30	81.82abc	2.74f
5	20	0.50	0.30	72.73abcd	3.58def
6	30	0.50	0.30	75.00abcd	4.24cdef
7	10	1.00	0.00	55.56de	4.80cd
8	20	1.00	0.00	80.00abc	4.22cdef
9	30	1.00	0.00	72.73abcd	4.43cde
10	10	1.00	0.30	63.64cd	4.26cdef
11	20	1.00	0.30	88.89ab	5.25c
12	30	1.00	0.30	63.64cd	2.84f
13	10	1.50	0.00	61.54cd	6.83ab
14	20	1.50	0.00	81.82abc	7.23a
15	30	1.50	0.00	70.00abcd	4.13cdef
16	10	1.50	0.30	25.00f	5.30bc
17	20	1.50	0.30	90.91a	4.08cdef
18	30	1.50	0.30	35.71ef	2.93ef
注：a ~ f表示生根率和平均根长指标下数据之间的显著性，带有相同字母表示差异不显著，反之则表示差异显著。Notes: a−f indicate the significance between the data under the rooting rate and average root length index, with the same letter indicates that the difference is not significant, and vice versa indicates that the difference is significant.

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[2]Liu M J. Chinese jujube: botany and horticulture[J]. Horticultural Review, 2006, 32: 229�?98. [3]何业�? 熊细�? 谢碧�? �? 枣树组织培养愈伤组织诱导的研究[J]. 中南林学院学�? 1997, 17(1): 13�?8.

He Y H, Xiong X M, Xie B X, et al. Inducement of calluses in the tissue culture of jujube trees[J]. Journal of Central South Forestry College, 1997, 17(1): 13�?8. [4]周瑞�? 刘孟�? 枣离体叶片高效再生植株的研究[J]. 园艺学报, 2006, 33(3): 625�?28. doi:10.3321/j.issn:0513-353X.2006.03.034

Zhou R J, Liu M J. Establishment of high-efficient in vitro leaf regeneration system in Chinese jujube[J]. Acta Horticulturae Sinica, 2006, 33(3): 625�?28. doi:10.3321/j.issn:0513-353X.2006.03.034 [5]李云, 王宇, 田砚�? �? 赞皇大枣叶片再生植株的初步研究[J]. 核农学报, 2003, 17(3): 187�?90. doi:10.3969/j.issn.1000-8551.2003.03.006

Li Y, Wang Y, Tian Y T, et al. A preliminary study on the plant regeneration from leaves of Zanhuang jujube[J]. Acta Agriculturae Nu-cleatae Sinica, 2003, 17(3): 187�?90. doi:10.3969/j.issn.1000-8551.2003.03.006 [6]胡影, 崔建�? 施满, �? 枣叶片愈伤组织的诱导和器官分化[J]. 甘肃林业科技, 1993, 18(4): 18�?9.

Hu Y, Cui J G, Shi M, et al. Induction and organ differentiation of calluses in jujube leaves[J]. Gansu Forestry Science and Technology, 1993, 18(4): 18�?9. [7]张春�? 黑果腺肋花楸离体叶片再生体系建立与原生质体分离培养研究[D]. 长春: 吉林农业大学, 2005.

Zhang C Y. Study on the establishment of ex vivo leaf regeneration system and protoplast isolation and culture of black fruit glandular rib rowan[D]. Changchun: Jilin Agricultural University, 2005. [8]李晓�? 张晓�? 灰枣试管苗移栽技术研究[J]. 河南农业科学, 2007, 36(8): 97�?8. doi:10.3969/j.issn.1004-3268.2007.08.032

Li X Q, Zhang X S. Research on transplanting technology of gray jujube test tube seedlings[J]. Journal of Henan Agricultural Sciences, 2007, 36(8): 97�?8. doi:10.3969/j.issn.1004-3268.2007.08.032 [9]陈宗�? 延志�? 齐龙. 枣叶片离体培养再生植株[J]. 植物生理学通讯, 1996, 32(1): 27�?8.

Chen Z L, Yan Z L, Qi L. Plantlet regeneration from leaves cultures of Ziziphus jujuba[J]. Plant Physiology Communications, 1996, 32(1): 27�?8. [10]孙清�? 孙洪�? 郑红�? �? 酸枣叶片不定植株的诱导[J]. 果树科学, 2000(1): 48�?1.

Sun Q R, Sun H Y, Zheng H J, et al. Study on the plant regeneration from leaves of sour jujube ( Zizyphus spinosusHu.)[J]. Journal of Fruit Science, 2000(1): 48�?1. [11]王玖�? 刘孟�? 代丽. 枣树组织培养研究进展[J]. 果树学报, 2002(5): 336�?39. doi:10.3969/j.issn.1009-9980.2002.05.011

Wang J R, Liu M J, Dai L. Advances in tissue culture of Chinese jujube[J]. Journal of Fruit Science, 2002(5): 336�?39. doi:10.3969/j.issn.1009-9980.2002.05.011 [12]陈宗�? 延志�? 薛皓, �? 沾化冬枣叶片培养和植株再生[J]. 植物生理学通讯, 2002, 38(6): 584.

Chen Z L, Yan Z L, Xue H, et al. In vitro culture of leaves and plantlet regeneration of Ziziphus jujubavar. z hanhua-dongzao[J]. Plant Physiology Communications, 2002, 38(6): 584. [13]胡桂�? 陈大�? 郑启�? �? 抗生素对台湾青枣茎段和愈伤组织生长的影响[J]. 华南农业大学学报, 2001, 22(2): 21�?3. doi:10.3969/j.issn.1001-411X.2001.02.007

Hu G B, Chen D C, Zheng Q F, et al. Effect of antibiotics on growth of stem segments and callus of ber ( Ziziphus mauritianaLam.)[J]. Journal of South China Agricultural University, 2001, 22(2): 21�?3. doi:10.3969/j.issn.1001-411X.2001.02.007 [14]何振�? 王玉�? 石武�? �? 山西特有品种梨枣叶片的组织培养及植株再生[J]. 植物生理学通讯, 2002, 38(5): 457.

He Z Y, Wang Y G, Shi W L, et al. Tissue culture and plant regeneration of leaves of Lizao endemic variety in Shanxi[J]. Plant Physiology Communications, 2002, 38(5): 457. [15]尹美�? 王玉�? 温银�? �? 梨枣叶片和茎段再生体系的建立[J]. 西北植物学报, 2005, 25(10): 1954�?959. doi:10.3321/j.issn:1000-4025.2005.10.005

Yin M Q, Wang Y G, Wen Y Y, et al. The construction of the regeneration system for the Chinese date cultivar Lizao to regenerate from its leaves and stems[J]. Acta Botanica Boreali-Occidentalia Sinica, 2005, 25(10): 1954�?959. doi:10.3321/j.issn:1000-4025.2005.10.005 [16]陈宗�? 张向�? 齐向�? �? 枣树叶片愈伤组织培养与再生植株的研究[J]. 福建林业科技, 2007, 34(3): 23�?7. doi:10.3969/j.issn.1002-7351.2007.03.006

Chen Z L, Zhang X Q, Qi X Y, et al. Callus culture from leaves and plant regeneration of jujube trees[J]. Journal of Fujian Forestry Science and Technology, 2007, 34(3): 23�?7. doi:10.3969/j.issn.1002-7351.2007.03.006 [17]王国�? 李晓�? 陈秋�? 枣不定芽再生体系的研究[J]. 中国农学通报, 2006, 22(4): 281�?83. doi:10.3969/j.issn.1000-6850.2006.04.074

Wang G P, Li X M, Chen Q F. Study on jujube adventitious bud regeneration system[J]. Chinese Agricultural Science Bulletin, 2006, 22(4): 281�?83. doi:10.3969/j.issn.1000-6850.2006.04.074 [18]黄健, 马锋�? 樊军�? �? 枣树离体叶片不定芽再生体系建立的研究[J]. 西北植物学报, 2006, 26(5): 942�?48. doi:10.3321/j.issn:1000-4025.2006.05.012

Huang J, Ma F W, Fan J F, et al. In vitro plant regeneration with adventitious buds of Zizyphus jujubaleaves[J]. Acta Botanica Boreali-Occidentalia Sinica, 2006, 26(5): 942�?48. doi:10.3321/j.issn:1000-4025.2006.05.012 [19]Ma C H, Ye X, Chen Y H. Anatomical observations of adventitious bud regenetation from leaf explants of Ziziphus jujubaMill. ‘Huizao’[J]. Horticulture Environment and Biotechnology, 2012, 53(4): 316�?19. doi:10.1007/s13580-012-0081-8 [20]潘青�? 白金, 王保�? 枣鲜食优质新品种‘京�?9’[J]. 中国果树, 2003(4): 3�?. doi:10.3969/j.issn.1000-8047.2003.04.002

Pan Q H, Bai J, Wang B Q. New high-quality jujube variety ‘Jingzao 39’[J]. China Fruits, 2003(4): 3�?. doi:10.3969/j.issn.1000-8047.2003.04.002 [21]程佑�? 安黎�? 浦铜�? �? 枣愈伤组织诱导和再生植株[J]. 西北植物学报, 2000, 20(3): 364�?69. doi:10.3321/j.issn:1000-4025.2000.03.008

Cheng Y F, An L Z, Pu T L, et al. Induction of callus and plant regeneration of Zizyphus jujubaMill.[J]. Acta Botanica Boreali-Occidentalia Sinica, 2000, 20(3): 364�?69. doi:10.3321/j.issn:1000-4025.2000.03.008 [22]王慧�? 马锋�? 张晓�? �? 鸡心枣试管苗叶片再生植株的研究[J]. 中国南方果树, 2007, 36(4): 48�?0. doi:10.3969/j.issn.1007-1431.2007.04.024

Wang H Y, Ma F W, Zhang X S, et al. Study on leaf regeneration plants of Jixinzao test tube seedlings[J]. South China Fruits, 2007, 36(4): 48�?0. doi:10.3969/j.issn.1007-1431.2007.04.024 [23]肖蓉, 王国�? 李春�? �? 辣椒枣试管苗叶片再生植株研究[J]. 山西农业科学, 2012, 40(3): 193�?96, 216. doi:10.3969/j.issn.1002-2481.2012.03.02

Xiao R, Wang G P, Li C Y, et al. Research of leaf regeneration plant in Ziziphus jujubaMill. Lajiaozao test tube seedlings[J]. Journal of Shanxi Agricultural Sciences, 2012, 40(3): 193�?96, 216. doi:10.3969/j.issn.1002-2481.2012.03.02 [24]周瑞�? �? Ziziphus jujubaMill.)离体叶片高效再生体系的建立[D]. 保定: 河北农业大学, 2004.

Zhou R J. Establishment of an efficient regeneration system for ex vivo leaves of Ziziphus jujubaMill.[D]. Baoding: Agricultural University of Hebei, 2004. [25]宁强, 刘晓�? 刘孟�? 冬枣叶片愈伤组织诱导研究[J]. 河北林果研究, 2008, 23(4): 348�?52. doi:10.3969/j.issn.1007-4961.2008.04.002

Ning Q, Liu X G, Liu M J. Callus inducement in tissue culture of jujube[J]. Hebei Forest Fruit Research, 2008, 23(4): 348�?52. doi:10.3969/j.issn.1007-4961.2008.04.002 [26]徐明, 代培�? 罗淑�? �? 哈密大枣叶片不定芽再生体系研究[J]. 新疆农业大学学报, 2014, 37(3): 185�?90. doi:10.3969/j.issn.1007-8614.2014.03.003

Xu M, Dai P H, Luo S P, et al. Study on adventitious bud regeneration system of Hami-Dazao leaves[J]. Journal of Xinjiang Agricultural University, 2014, 37(3): 185�?90. doi:10.3969/j.issn.1007-8614.2014.03.003 [27]李静, 张景�? 郭伟, �? 毛白杨叶片愈伤组织的诱导、芽的分化及不定根的再生[J]. 山东林业科技, 2007, 168(1): 52�?3. doi:10.3969/j.issn.1002-2724.2007.01.020

Li J, Zhang J C, Guo W, et al. Callus induction, bud differentiation and adventitious root regeneration from leaves of Populus tomentosa[J]. Shandong Forestry Science and Technology, 2007, 168(1): 52�?3. doi:10.3969/j.issn.1002-2724.2007.01.020 [28]刘晓�? 单梅�? 苏帅, �? 山杏组培苗生根培养基优化及移栽[J]. 分子植物育种, 2020, 18(6): 1999�?005. doi:10.13271/j.mpb.018.001999

Liu X G, Shan M H, Su S, et al. Transplanting and optimization of rooting medium for apricot tissue cultured seedlings[J]. Molecular Plant Breeding, 2020, 18(6): 1999�?005. doi:10.13271/j.mpb.018.001999