豌豆愈伤组织及不定芽诱导影响因素的研究

doi:10.16035/j.issn.1001-7283.2016.03.007

摘要/Abstract

摘要：

以豌豆品种陇豌1号为试验材料,对愈伤组织和不定芽诱导过程中种子处理、外植体选择、最适培养基、不定芽诱导及生根等因素进行研究。确定了最适的种子消毒处理为75%乙醇消毒30s,0.1%升汞消毒10min。以下胚轴为外植体诱导愈伤组织,最适激素配比为0.2mg/L 2,4-D和1.0mg/L TDZ,愈伤组织诱导率为91.11%。具有分化能力的愈伤组织持续培养在只添加TDZ的培养基上即可诱导出不定芽,但这些不定芽生根率仅为8.89%,为此分析了影响生根的因素,为后续的研究提供依据。

关键词: 豌豆, 愈伤组织, 不定芽

Abstract:

The pea variety Longwan No.1 was used as experimental material to study the factors in callus and adventitious bud induction process, such as seed treatment,explants selection, optimal medium, adventitious bud and rooted inducing.The results showed that the optimal disinfection treatments for seed was 75% alcohol solution for 30 seconds, and then 0.1% HgCl₂ for 10min. Hypocotyl as the explants, the optimal culture medium of callus was 0.2mg/L 2,4-D and 1.0mg/L TDZ with an induction rate of 91.11%. Only the addition of TDZ could induce adventitious bud,but the rooting rate was only 8.89%. We also analyzed the factors affecting the growth of root,which provided the basis for subsequent research.

Key words: Pea, Callus, Adventitious shoot

张丽娟,陆建英,王昶,闵庚梅,杨晓明. 豌豆愈伤组织及不定芽诱导影响因素的研究[J]. 作物杂志, 2016 (3): 33–36

Lijuan Zhang,Jianying Lu,Chang Wang,Gengmei Min,Xiaoming Yang. The Study on the Influence Factors of Callus and Adventitious Bud Induction in Pea[J]. Crops, 2016(3): 33–36

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参考文献 9

[1]	贺晨帮, 宗绪晓 . 豌豆种质资源形态标记遗传多样性分析. 植物遗传资源学报, 2011,12(1):42-48. doi: 10.3969/j.issn.1005-9393.2009.05.006
[2]	Das A, Kumar S, Nandeesha P , et al. An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via.shoot organogenesis. Journal of Plant Biochemistry and Biotechnology, 2014,23(2):184-189. doi: 10.1007/s13562-013-0200-3
[3]	Malmberg R L . Regeneration of whole plants from callus culture of diverse genetic lines of Pisum sativum L. Planta. 1979,146(2):243-244. doi: 10.1007/BF00388239 pmid: 24318066
[4]	Popiers D, Flandre F ,Sangwan-Norreel B S.Intensification de la regeneration du pois (Pisum sativum L.),par le thidiazuron,via la formation de structures caulinaires organogenes. Canadian Journal Botany, 1997,75(3):492-500. doi: 10.1139/b97-053
[5]	Tzitzikas E N, Bergervoet M, Raemakers K , et al. Regeneration of Pea (Pisum sativum L.)by a cyclic organogenic system. Plant Cell Reports, 2004,23(7):453-461. doi: 10.1007/s00299-004-0865-0 pmid: 15372196
[6]	Murthy B N S, Murch S, Saxena P K . Thidiazuron:A potent regulator of in vitro plant morphogenesis. In Vitro Cellular and Developmental Biology, 1998,34(4):267-275. doi: 10.1007/BF02822732
[7]	苏承刚, 吴学科, 郑占伟 , 等. 食荚型豌豆组织培养和植株再生研究. 西南师范大学学报(自然科学版), 2007,32(4):30-32. doi: 10.3969/j.issn.1000-5471.2007.04.007
[8]	Zhihui S, Tzitzikas M, Raemakers K , et al. Effect of TDZ on plant regeneration from mature seeds in pea (Pisum sativum). In Vitro Cellular & Developmental Biology—Plant, 2009,45(6):776-782. doi: 10.1007/s11627-009-9212-z
[9]	Petra B, Beate M, Hans-Jörg J . Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast derived pea callus. Plant Cell Reports, 1995,15(1):26-29. doi: 10.1007/BF01690247

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处理 Treatments	75%乙醇消毒时间(s) Disinfection time of 75% alcohol solution	0.1%升汞消毒时间(min) Disinfection time of0.1% HgCl₂	出苗率(%) Emergence rate	污染情况 Condition of contamination
C1	30	0	14.44±1.11g	+++
C2	30	5	54.45±2.22e	++
C3	30	10	92.22±2.94a	-
C4	30	15	93.33±1.93a	-
C5	30	20	84.44±1.11b	-
C6	30	30	51.11±1.11e	-
C7	60	0	27.78±2.94f	+++
C8	60	5	70.00±1.92d	+
C9	60	10	90.00±1.92ab	-
C10	60	15	87.78±1.11ab	-
C11	60	20	76.67±1.93c	-
C12	60	30	48.89±1.11e	-

外植体Explants	接种外植体数No.of explants(个)	产生愈伤组织外植体数No.of Callus induction(个)	愈伤组织诱导率Callus induction rate(%)
下胚轴Hypocotyl	90×3	224	82.97±1.34aA
子叶Cotyledon	90×3	64	23.70±0.74bB
胚根Radicel	90×3	17	6.30±1.34cC

组合代号 Code of combination	2,4-D浓度(mg/L) 2,4-D concentration	6-BA浓度(mg/L) 6-BA concentration	TDZ浓度(mg/L) TDZ concentration	愈伤组织诱导率(%) Callus induction rate
Y1	0.2	0	0	0.00±0.00e
Y2	1.0	0	0	0.00±0.00e
Y3	0.2	0.2	0	57.78±2.94d
Y4	0.2	0.5	0	74.44±2.94c
Y5	0.2	0	0.5	83.33±1.93b
Y6	0.2	0	1.0	91.11±1.11a

TDZ浓度(mg/L) TDZ concentration	接种愈伤组织块数 No.of callus	形成不定芽的愈伤组织块数 No.of callus with shoots	不定芽分化率(%) Shoot differentiation rate	不定芽增殖数(个) No.of shoots per explant
0.5	30×3	75	83.33±3.85a	10.00±1.15c
1.0	30×3	81	90.00±1.92a	17.67±1.20c
2.0	30×3	82	91.11±2.94a	25.00±2.52b
4.0	30×3	82	91.11±1.11a	26.33±0.88a

激素浓度(mg/L) Hormone concentration	生根率 Rooting rate(%)
激素浓度(mg/L) Hormone concentration	NAA	IBA	IAA
0.5	0	0	0
1.0	0	0	0
2.0	0	8.89	0
4.0	0	0	0