菜薹和芥蓝种间杂交胚挽救技术研究

吕凤仙; 李崇娟; 杨鼎; 胡靖锋; 杨红丽; 兰梅; 徐学忠; 张丽琴; 和江明; 董相书

doi:10.19303/j.issn.1008-0384.2021.11.005

菜薹和芥蓝种间杂交胚挽救技术研究

doi: 10.19303/j.issn.1008-0384.2021.11.005

吕凤仙^{1, 2,},
李崇娟^{1, 2},
杨鼎^{1, 2},
胡靖锋^{2, 3},
杨红丽^{2, 3},
兰梅^{2, 3},
徐学忠^{2, 3},
张丽琴^{2, 3},
和江明^{2, 3, ,},
董相书¹

1.
云南大学, 云南昆明　650504
2.
云南省农业科学院园艺作物研究所, 云南　昆明　650205
3.
国家蔬菜改良分中心云南分中心, 云南　昆明　650205

基金项目: 云南省“万人计划”产业领军人才项目（YNWR-CYJS-2018-040）；国家大宗蔬菜产业技术体系昆明综合试验站项目（CARS-23-G37）；云南省科技计划重大科技专项（2019ZG001）

详细信息

作者简介:
吕凤仙（1997−），女，硕士生，研究方向：十字花科蔬菜育种（E-mail：2547682199@qq.com）

通讯作者:
和江明（1969−），男，研究员，研究方向：十字花科作物生物技术育种（E-mail：hejiangming666@qq.com.cn）

中图分类号: S 634
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- 被引次数: 0
出版历程
- 收稿日期: 2021-05-31
- 修回日期: 2021-10-14
- 网络出版日期: 2021-12-30
- 刊出日期: 2021-11-28

Embryo Rescue of Interspecific Brassica campestris x B. albograbra Hybrids

LYU Fengxian^{1, 2
,},
LI Chongjuan^{1, 2},
YANG Ding^{1, 2},
HU Jingfeng^{2, 3},
YANG Hongli^{2, 3},
LAN Mei^{2, 3},
XU Xuezhong^{2, 3},
ZHANG Liqin^{2, 3},
HE Jiangming^{2, 3
, ,},
DONG Xiangshu¹

1.
Yunnan University, Kunming, Yunnan　650504, China
2.
Horticultural Crops Research Institute, Yunnan Academy Agricultural Science, Kunming, Yunnan　650205, China
3.
Yunnan Branch of National Vegetable Improvement Center, Kunming, Yunnan　650205, China

摘要

摘要: 目的获得适合菜薹和芥蓝种间杂种的胚挽救实验方案。方法以菜薹（菜心）作为母本，芥蓝作为父本，进行种间杂交，对种间杂种进行人工胚挽救培养，设置8种不同成分的胚挽救培养基，6个不同取材时间，4种增殖培养基，9种不同成分生根培养基。结果 ERM₄（MS+6-BA 0.5 mg·L⁻¹+NAA 0.5 mg·L⁻¹+KT 1.0 mg·L⁻¹+AC 1.0 g·L⁻¹）出胚率达26.32%，为最佳胚挽救培养基；授粉后第12天取材，出胚率达26.32%，为最适宜取材时间；DM₁（MS+6-BA 0.9 mg·L⁻¹+NAA 0.1 mg·L⁻¹）培养基增殖系数最大，为5.37，为最佳增殖培养基；RM_4-2（1/2MS+IBA0.5 mg·L⁻¹）培养基生根率为37.21%，平均生根时间为8.33 d，生根最快，为最佳生根培养基。结论菜薹与芥蓝种间杂交，于授粉后12 d取材，胚珠接种在ERM₄培养基上，出胚后接种在DM₁培养基上增殖，在RM_4-2培养基上生根，能够显著提高种间杂种的获得率。
- 菜薹 /
- 芥蓝 /
- 种间杂交 /
- 胚挽救 /
- 增殖 /
- 不定根诱导
Abstract: Objective Methods suitable for embryo rescue in the interspecific hybridization between Brassica campestris and B. albograbra were investigated. Method The interspecific crossing was carried out using B. campestris L. as female parent and B. albograbra L. H. Bailey as male parent. An embryo rescue culture for the hybridization was conducted with 8 different media at 6 sampling times followed subsequently by 4 proliferation media and 9 rooting media. Result The embryo genesis rate at 26.32% of ERM₄ medium consisting of MS, 0.5mg 6-BA·L⁻¹, 0.5mg NAA·L⁻¹, 1.0mg KT·L⁻¹, and 1.0g AC·L⁻¹ was the highest among all. The samples collected 12d after pollination had the best rate of 26.32%, DM₁ (i.e., MS+ 0.9mg 6-BA·L⁻¹+0.1mg NAA·L⁻¹) the greatest proliferation coefficient of 5.37, and RM_4-2 (i.e., 1/2MS+ 0.5mg IBA·L⁻¹) the top rate of 37.21% and average 8.33d on rooting. Conclusion The optimum conditions for the embryo rescue in the interspecific hybridization between B. campestris L. and B. albograbra L. H. Bailey included sampling at 12d after pollination and following by culture on ERM₄ medium for embryo induction, on DM₁ for ovule proliferation, and on RM_4-2 for seedling rooting.
- Brassica campestris L. /
- Brassica albograbra L. H. Bailey /
- interspecies cross /
- embryo rescue /
- proliferation /
- adventitious root induction

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图 1 菜薹×芥蓝种间杂种胚挽救情况

注：A，胚珠萌发；B，胚挽救幼苗；C，成活胚增殖分化

Figure 1. Embryo rescue of interspecific B. campestris × B. albograbra hybrids

Note: A: Ovule germination; B: embryo rescued seedling; C: proliferation and differentiation of survival embryos.

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图 2 菜薹×芥蓝种间杂种生根过程不同生长素生长果

注：A：NAA生根效果；B：IBA生根效果；C：IAA生根效果；D：未加激素MS培养基生根效果

Figure 2. Effects of auxins on growth during rooting of interspecific B. campestris × B. albograbra hybrids

Note: A: Rooting affected by NAA; B: rooting affected by IBA; C: rooting affected by IAA; D: seedling rooting on MS without hormone addition.

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图 3 菜薹×芥蓝种间杂种生根过程盐害现象

Figure 3. Damage induced by salt at rooting stage of interspecific B. campestris × B. albograbra hybrids

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图 4 菜薹×芥蓝种间杂种生根过程

Figure 4. Rooting of interspecific B. campestris × B. albograbra hybrids

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图 5 菜薹×芥蓝种间杂种植株形态

注：A，菜薹；B、C，种间杂种；D，芥蓝

Figure 5. Morphology of interspecific B. campestris × B. albograbra hybrid plant

Note：A: B. campestris; B and C: interspecific hybrid; D: B. albograbra.

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图 6 菜薹×芥蓝种间杂种花器官形态

注：A：F菜薹；B：E种间杂种；C：D芥蓝

Figure 6. Morphology of floral organ of interspecific B. campestris × B. albograbra hybrid

Note: A and F: B. campestris; B and E: interspecific hybrid; C and D: B. albograbra.

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表 1 菜薹×芥蓝种间杂种胚挽救培养基成分

Table 1. Components in embryo rescue medium for interspecific B. campestris × B. albograbra hybrids

编号 No.	基本培养基 Minimal medium	6-BA质量浓度 6-BA mass concentration/ （mg·L⁻¹）	NAA质量浓度 NAA mass concentration/ （mg·L⁻¹）	KT质量浓度 KT mass concentration/ （mg·L⁻¹）	AC含量 AC content/ （g·L⁻¹）
ERM₁	MS	0.00	0.00	0.00	1.00
ERM₂	MS	0.30	0.00	0.00	1.00
ERM₃	MS	0.50	0.00	0.00	1.00
ERM₄	MS	0.50	0.50	1.00	1.00
ERM₅	MS	1.50	0.50	1.00	1.00
ERM₆	MS	3.00	0.50	1.00	1.00
ERM₇	MS	4.50	0.50	1.00	1.00
ERM₈	MS	5.00	0.50	1.00	1.00

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表 2 菜薹×芥蓝种间杂种增殖培养基成分

Table 2. Components in propagation medium for interspecific B. campestris × B. albograbra hybrids

编号 No.	基本培养基 Minimal medium	6-BA质量浓度 6-BA mass concentration/ （mg·L⁻¹）	NAA质量浓度 NAA mass concentration/ （mg·L⁻¹）
DM₁	MS	0.90	0.10
DM₂	1/2MS	0.90	0.10
DM₃	MS	0.50	0.01
DM₄	1/2MS	0.50	0.01

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表 3 菜薹×芥蓝种间杂种生根培养基成分

Table 3. Components in rooting medium for interspecific B. campestris × B. albograbra hybrids

编号　 No.	基本培养基 Minimal medium	NAA质量浓度 NAA mass concentration/ （mg·L⁻¹）	IBA质量浓度 IBA mass concentration/ （mg·L⁻¹）	IAA质量浓度 IAA mass concentration/ （mg·L⁻¹）
RM_1-1	MS	0.20	0.00	0.00
RM_1-2		0.50	0.00	0.00
RM_2-1		0.00	0.20	0.00
RM_2-2		0.00	0.50	0.00
RM_3-1		0.00	0.00	0.20
RM_3-2		0.00	0.00	0.50
RM_4-1	1/2MS	0.20	0.00	0.00
RM_4-2		0.00	0.50	0.00
RM_4-3		0.00	0.00	0.50

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表 4 菜薹×芥蓝杂种胚珠在不同培养基上出胚率

Table 4. Embryogenesis rate of B. campestris × B. albograbra hybrid ovules on different media

编号 No.	胚珠培养数 Number of ovules	成胚数 Number of embryogenesis	出胚率 Embryogenesis rate/%
ERM₁	105	0	0.00±0.00 f
ERM₂	75	1	1.33±0.02 ef
ERM₃	78	2	2.56±1.25 e
ERM₄	95	25	26.32±0.78 a
ERM₅	88	16	18.18±0.93 b
ERM₆	85	13	15.29±1.00 c
ERM₇	101	10	9.90±0.89 d
ERM₈	103	8	7.77±0.90 d
注：不同小写字母表示差异达0.05显著水平。下同。 Note: Different lowercase letters indicate significant difference at P<0.05. Same for the following.

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表 5 菜薹×芥蓝不同取材时间杂种胚珠出胚率

Table 5. Embryogenesis rate of B. campestris × B. albograbra hybrid ovules with differed sampling time

授粉后天数 Days after pollination/d	胚珠培养数 Number of ovules	出胚数 Number of embryogenesis	出胚率 Embryogenesis rate/%
10	90	4	4.44±1.93 b
11	88	9	10.23±1.97 b
12	95	25	26.32±2.77 a
13	382	40	10.47±0.92 b
14	150	10	6.67±3.34 b
15	60	2	3.33±2.89 c

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表 6 菜薹×芥蓝种间杂种成活胚珠在不同培养基上增殖分化情况

Table 6. Proliferation and differentiation of viable ovules in B. campestris × B. albograbra hybrids on different media

编号 No.	原有芽数 Number of original bud	出芽数 Number of budding	增殖系数 Multiplication coefficient
DM₁	79	424	5.37±0.05 a
DM₂	76	306	4.02±0.38 b
DM₃	80	332	4.15±0.25 b
DM₄	78	226	2.90±0.15 c

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表 7 菜薹×芥蓝种间杂种不同培养基生根情况

Table 7. Rooting of interspecific B. campestris × B. albograbra hybrids on different media

编号 No.	培养幼苗数 Number of cultured seedlings	生根幼苗数 Number of rooting seedlings	生根率 Rate of rooting/%	平均生根时间 Mean rooting time/d	移栽后成活幼苗 Number of survived seedlings after transplanting	移栽成活率 Rate of transplanting survival/%
RM_1-1	27	11	40.74±2.20 ab	11.00±1.00 b	11	100
RM_1-2	28	4	14.29±3.07 e	11.33±0.58 b	4	100
RM_2-1	24	6	25.00±3.13 cd	8.67±0.58 b	6	100
RM_2-2	25	9	36.00±2.57 bc	8.67±0.58 b	9	100
RM_3-1	27	5	18.52±3.03 d	14.33±1.53 a	5	100
RM_3-2	27	7	25.93±2.75 c	15.00±1.00 a	7	100
RM_4-1	45	20	44.44±1.93 a	11.33±0.58 b	20	100
RM_4-2	43	16	37.21±1.47 cd	8.33±0.58 c	16	100
RM_4-3	47	13	27.66±1.54 c	14.67±1.53 a	13	100

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