郁金樱愈伤组织诱导及植株再生

doi:10.11983/CBB20205

摘要/Abstract

摘要： 为建立郁金樱(Cerasus serrulata var. lannesiana cv. ‘Grandiflora’)再生体系, 以多年生母株小叶、一年生嫁接苗小叶、腋芽诱导小叶和增殖一代小叶为外植体, 探讨不同外植体和植物激素组合对郁金樱愈伤组织诱导、不定芽分化、增殖和生根的影响。结果表明, 4种外植体均可诱导出愈伤组织, 除多年生母株小叶外皆分化出不定芽, 外植体幼化程度越高, 后期培养潜力越大。以增殖一代小叶为外植体效果最佳, 其最适愈伤组织诱导培养基为MS+0.5 mg·L^-16-BA+1.0 mg·L^-12,4-D, 诱导率达96.22%; 最佳分化培养基为MS+1.0 mg·L^-16-BA+0.1 mg·L^-12,4-D+0.1 mg·L^-1TDZ, 分化率达78.14%; 最佳增殖培养基为MS+1.0 mg·L^-16-BA, 增殖系数可达7.85; 最佳生根培养基为不含任何激素的1/2MS培养基, 生根率达100%。不同外植体获得的再生植株移栽成活后生长差异显著, 以增殖一代小叶诱导的再生植株长势最佳。

关键词: 郁金樱, 愈伤组织, 外植体, 植株再生, 组织培养

Abstract: In order to establish the regeneration system of Cerasus serrulata var. lannesiana cv. ‘Grandiflora’, the effects of different explants and plant hormone combinations on callus induction, adventitious bud differentiation, proliferation and rooting were studied using perennial mother plant leaflets, annual grafted seedling leaflets, axillary bud induction leaflets and proliferative first-generation leaflets as explants. The results showed that callus could be induced from all four explants, and adventitious buds could be derived from all explants except the leaflets of perennial mother plants. The higher the degree of explants’ juvenility, the greater the success of subsequent culture, with the best explants were the proliferative first generation leaflets. The best medium for callus induction was MS+0.5 mg·L^-1 6-BA+1.0 mg·L^-1 2,4-D, and the induction rate was 96.22%. The optimal medium for differentiation was MS+1.0 mg·L^-1 6-BA+0.1 mg·L^-1 2,4-D+0.1 mg·L^-1 TDZ, and the differentiation rate was 78.14%. The optimal medium for proliferation was MS+1.0 mg·L^-1 6-BA, and the proliferation coefficient reached 7.85. The optimal medium for rooting was 1/2MS without any hormone, and the regenerated plants with 100% rooting rate were obtained. The regenerated plants from different explants grew very differently, and those induced from first generation of proliferative leaflets had the best growth.

Key words: Cerasus serrulata var. lannesiana cv. ‘Grandiflora’, callus, explants, plant regeneration, tissue culture

罗钱, 张燕莎, 欧静. 郁金樱愈伤组织诱导及植株再生. 植物学报, 2021, 56(4): 451-461.

Qian Luo, Yansha Zhang, Jing Ou. Callus Induction and Plant Regeneration of Cerasus serrulata var. lannesiana cv. ‘Grandiflora’. Chinese Bulletin of Botany, 2021, 56(4): 451-461.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB20205

https://www.chinbullbotany.com/CN/Y2021/V56/I4/451

图/表 10

表1 郁金樱愈伤组织分化不同植物激素正交设计

Table 1 Orthogonal design of different plant hormones in callus differentiation of Cerasus serrulata

Concentrations of plant hormones (mg·L^-1)	Treatment code
Concentrations of plant hormones (mg·L^-1)	1	2	3	4	5	6	7	8	9
6-BA	0.5	0.5	0.5	1	1	1	2	2	2
2,4-D	0.1	0.5	1	0.1	0.5	1	0.1	0.5	1
TDZ	0.01	0.1	0.5	0.1	0.5	0.01	0.5	0.01	0.1

图1 郁金樱不同外植体愈伤组织的平均诱导率不同小写字母表示不同处理间差异显著(P<0.05)。

Figure 1 Average callus induction rates in different explants of Cerasus serrulata Different lowercase letters indicate significant differences among different treatments (P<0.05).

图2 郁金樱愈伤组织诱导、分化及增殖 (A) 腋芽诱导小叶; (B1)-(B3) 多年生母株小叶愈伤组织诱导及分化; (C1)-(C7) 一年生嫁接苗小叶愈伤组织诱导、不定芽分化及增殖过程; (D1)-(D7) 腋芽诱导小叶的愈伤组织诱导、不定芽分化及增殖过程; (E1)-(E7) 增殖一代小叶的愈伤组织诱导、不定芽分化及增殖过程。Bars=1 cm

Figure 2 Callus induction, differentiation and proliferation of Cerasus serrulata (A) Axillary buds induce leaflets; (B1)-(B3) Leaflet callus induction and differentiation of perennial mother plant; (C1)-(C7) Leaflet callus induction, adventitious bud differentiation and proliferation of one-year grafted seedlings; (D1)-(D7) The process of callus induction, adventitious bud differentiation and proliferation induced from axillary buds; (E1)-(E7) The process of callus induction, adventitious bud differentiation and proliferation of the proliferative generation of leaflets. Bars=1 cm

表2 不同激素组合及外植体对郁金樱愈伤组织诱导的影响(平均值±标准误)

Table 2 Effects of different hormone combinations and explants on Cerasus serrulata callus induction (means±SE)

No.	6-BA (mg·L^-1)	2,4-D (mg·L^-1)	Callus induction rate (%)
No.	6-BA (mg·L^-1)	2,4-D (mg·L^-1)	Leaflet of perennial mother plant	Annual grafted seedling leaflet	Axillary buds induce leaflets	Proliferate one generation of leaflets
1	0	0.5	0 l	42.87±1.51 i	66.09±0.72 g	66.51±0.33 i
2	0	1.0	23.91±0.51 j	49.32±1.01 h	68.46±1.00 ef	72.86±0.59 g
3	0	1.5	46.28±0.58 h	58.92±0.72 f	67.64±0.80 f	76.23±0.96 ef
4	0	2.0	57.35±0.68 e	68.63±0.60 d	72.52±1.16 d	80.77±0.59 d
5	0	3.0	57.15±0.32 e	61.31±1.34 e	69.61±0.67 e	77.63±0.71 e
6	0.5	0.5	16.68±1.45 k	55.08±0.82 g	63.67±0.54 h	71.12±0.20 h
7	0.5	1.0	60.09±0.98 d	61.43±1.40 e	69.04±0.70 ef	96.22±0.94 a
8	0.5	1.5	49.31±0.70 g	74.11±0.78 c	83.07±0.73 c	94.63±0.42 b
9	0.5	2.0	68.55±1.04 a	80.71±0.49 a	88.74±0.45 a	91.40±0.80 c
10	0.5	3.0	62.52±1.09 c	77.62±1.11 b	86.17±0.72 b	77.05±0.77 ef
11	1	0.5	27.85±0.69 i	62.06±0.73 e	68.09±0.23 f	76.63±0.82 ef
12	1	1.0	45.64±0.99 h	69.25±0.42 d	73.28±0.64 d	81.36±1.09 d
13	1	1.5	51.03±1.17 f	73.81±0.92 c	83.64±1.06 c	92.18±2.00 c
14	1	2.0	65.46±0.61 b	60.95±0.30 e	67.87±0.98 f	75.57±0.90 f
15	1	3.0	59.89±1.22 d	54.64±1.00 g	65.07±0.82 f	73.85±0.63 g

图3 不同植物激素对郁金樱不同外植体愈伤组织分化的影响分化培养基编号1-9同表1。不同小写字母表示不同处理间差异显著(P<0.05)。

Figure 3 Effects of different plant hormones on Cerasus serrulata callus differentiation in different explants Differentiation medium No. 1-9 are the same as Table 1. Different lowercase letters indicate significant differences at P<0.05 among different treatments.

表3 不同植物激素对郁金樱不同外植体愈伤组织分化的方差分析

Table 3 Variance analysis of Cerasus serrulata callus differentiation of different explants with different plant hormones

Different explants	Source	df	Sum of squares	Mean square	F	P
Annual grafted seedling leaflet	6-BA	2	2622.375	1311.187	77.591	0.000**
	2,4-D	2	57.819	28.910	1.711	0.206
	TDZ	2	128.789	64.394	3.811	0.040*
	Error	20	337.975	16.899	-	-
	Sun	27	13723.723	-	-	-
Axillary buds induce leaflets	6-BA	2	1476.646	738.323	42.619	0.000**
	2,4-D	2	144.656	72.328	4.175	0.031*
	TDZ	2	19.924	9.962	0.575	0.572
	Error	20	346.474	17.324	-	-
	Sun	27	29649.142	-	-	-
Proliferate one generation of leaflets	6-BA	2	3360.867	1680.433	29.026	0.000**
	2,4-D	2	600.965	300.483	5.19	0.015*
	TDZ	2	182.009	91.005	1.572	0.232
	Error	20	1157.866	57.893	-	-
	Sun	27	79113.749	-	-	-

表4 不同植物激素对郁金樱外植体不定芽增殖的影响(平均值±标准误)

Table 4 Effects of different plant hormones on Cerasus serrulata adventitious bud proliferation of different explants (means±SE)

No.	6-BA (mg·L^-1)	NAA (mg·L^-1)	Proliferation coefficient
No.	6-BA (mg·L^-1)	NAA (mg·L^-1)	Annual grafted seedling leaflets	Axillary buds induce leaflets	Proliferate one generation of leaflets
1	0.5	0	0 e	2.42±0.04 g	3.32±0.46 e
2	0.5	0.1	0.88±0.77 d	3.52±0.07 f	2.70±0.24 f
3	0.5	0.5	2.17±0.27 c	2.39±0.05 g	2.91±0.29 ef
4	1	0	2.38±0.04 c	4.46±0.12 e	7.85±0.33 a
5	1	0.1	2.17±0.62 c	4.99±0.11 c	5.57±0.20 cd
6	1	0.5	3.13±0.05 ab	4.64±0.08 d	6.30±0.14 b
7	2	0	2.61±0.07 bc	5.39±0.05 a	5.26±0.27 d
8	2	0.1	3.07±0.06 b	5.18±0.19 b	5.82±0.19 c
9	2	0.5	3.53±0.04 a	4.73±0.08 d	3.38±0.15 e
Average			2.22±1.12 C	4.19±1.10 B	4.79±1.73 A

表5 不同植物激素对郁金樱生根的影响(平均值±标准误)

Table 5 Effects of plant hormones on rooting of Cerasus serrulata (means±SE)

IBA (mg·L^-1)	NAA (mg·L^-1)	Rooting rate (%)	Number of roots (strips)
0	0	100 a	12.82±0.54 a
0	0.5	86.72±1.62 b	9.24±1.24 b
0	1	0 e	0 e
0.5	0	68.32±2.95 c	5.64±0.26 c
0.5	0.5	44.09±1.74 d	3.24±0.48 d

图4 郁金樱不同外植体诱导再生植株生根及移载 (A) 不定根原始体突起; (B) 肉质主根; (C) 主根上长出侧根; (D) 直接长出须根; (E)-(G) 一年生嫁接苗小叶植株再生、生根及移栽; (H)-(J) 腋芽诱导小叶再生植株生根及移栽; (K)-(M) 增殖一代小叶再生植株生根及移栽。Bars=2 cm

Figure 4 Rooting and transplantation of regenerated plants induced by different explants of Cerasus serrulata (A) Adventitious roots protuberant; (B) Fleshy taproots; (C) Lateral roots on the taproot; (D) Directly grow fibrous roots; (E)-(G) Leaflet regeneration of annual grafted seedlings, rooting and transplanting; (H)-(J) The axillary buds induced the rooting and transplanting of regenerated leaflets; (K)-(M) Proliferation of a generation of regenerated leaflets, rooting and transplanting. Bars=2 cm

图5 郁金樱不同外植体诱导的再生植株的生长状态

Figure 5 The growth status of regenerated plants induced by different explants of Cerasus serrulata

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