Somatic embryogenesis and plantlet regeneration in <i>Pinus elliottii</i>and its hybrids

HU Jiwen; GUO Wenbing; DENG Leping; ZHONG Suiying; WANG Weimin; ZHAO Fencheng; HUANG Ting; WU Huishan; LI Yiliang; LIAO Fangyan

doi:10.7671/j.issn.1001-411X.201804001

Journal of South China Agricultural University > 2019 > 40(1): 107-115. > DOI: 10.7671/j.issn.1001-411X.201804001

HU Jiwen, GUO Wenbing, DENG Leping, et al. Somatic embryogenesis and plantlet regeneration in Pinus elliottiiand its hybrids[J]. Journal of South China Agricultural University, 2019, 40(1): 107-115. DOI: 10.7671/j.issn.1001-411X.201804001

Citation:

PDF (998 KB)

Somatic embryogenesis and plantlet regeneration in Pinus elliottiiand its hybrids

1.
Guangdong Provincial Key Laboratory of Silviculture Protection and Utilization/Guangdong Academy of Forestry, Guangzhou 510520, China
2.
Taishan Hongling Seed Orchard, Jiangmen 529223, China

More Information

Received Date: March 31, 2018
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
To define the conditions suitable for somatic embryogenesis of Pinus elliottii and its hybrids, and develop a mature protocol for somatic embryogenesis maturation and germination.

Method
The immature zygotic embryos (with endosperm) of two P. elliottii families (EE1, EE2) and two P. elliottii hybrid (EC, EH) were sampled in June 2016. Three series of medium from induction, proliferation maturation to germination were set to compare the effects of explant collection date, family and basal medium on embryogenic callus induction. The embryogenic callus were identified through microscopic observation, and choosen for proliferation, maturation and germination culture. Finally the regenerated plantlets were developed.

Result
The zygotic embryo development processes of P. elliottii and hybrids were divided into eight stages. The immature embryos at stage Ⅱ and Ⅲ were suitable for somatic embryogenesis. The embryos at stage Ⅲ first appeared in EC. The embryogenic callus with embryonal suspensor mass (ESM) could be further proliferated. The induction mediums (T1, T2 and T3) played vital roles in callus formation and embryogenic callus proportion. T1 medium had the highest callus induction efficiency (49.0%), and T2 medium had the highest embryogenic callus proportion (22.4%).The mediums had genotype-specific induction rates. The overall induction rate of T1 medium was low, and T2 medium had the highest induction rate (5.82%) for EE1. T3 medium was suitable for all tested materials, with the highest mean induction rate of 3.75%. The somatic embryo induction rates of EE1 and EH gradually increased with the sampling time extension, while the somatic embryo induction rates of EE2 and EC gradually decreased. The results of somatic embryo induction rate were basically corresponded with zygote development stages. The stage Ⅲ zygotic embryo appeared late, and its induction rate was low in early stage. After the embryogenic callus were subcultured for 24 times, the embryo activities progressively decreased. The maturation medium T1S and T3S could accomplish embryo maturation. Their average ripening efficiencies in per gram of mature medium were 23.3 and 15.9 cotyledons, respectively. The germination rate was 32.1% and the transplanting rate was 47.8%.

Conclusion
All tested materials can induce somatic embryo in T3 medium. T3 medium is widely applicable, and all materials have high induction rates when zygotic embryo appears at stage Ⅲ, illustrating that stage Ⅲ may be the optimal for somatic embryo induction. The experiment establishes the protocol for somatic embryogenesis and plantlet regeneration.
- Pinus elliottii,
- zygotic embryo,
- somatic embryogenesis,
- plant regeneration,
- hybridization,
- embryogeniccallus,
- inductionmedium

FullText(HTML)

References (34)

References

[1]	李彦杰, 栾启福, 沈丹玉, 等. 湿地松自由授粉家系松脂组分遗传变异研究[J]. 林业科学研究, 2012, 25(6): 773-779. doi: 10.3969/j.issn.1001-1498.2012.06.016
[2]	WEN X, KUANG Y, SHI M, et al. Biology of Hylobitelus xiaoi(Coleoptera: Curculionidae), a new pest of slash pine, Pinus elliottii[J]. J Econ Entomol, 2004, 97(6): 1958-1964. doi: 10.1093/jee/97.6.1958
[3]	MEANS D B, PALIS J G, BAGGETT M. Effects of slash pine silviculture on a florida population of flatwoods salamander[J]. Conserv Biol, 1996, 10(2): 426-437. doi: 10.1046/j.1523-1739.1996.10020426.x
[4]	张帅楠. 湿地松材性测定技术研究与多性状联合选择[D]. 北京: 中国林业科学研究院, 2017.
[5]	宗亦臣, 郑勇奇, 马锡权, 等. 湿加松良种‘中林1号’[J]. 湖南林业科技, 2017, 44(1): 76-77. doi: 10.3969/j.issn.1003-5710.2017.01.015
[6]	吴丽君. 湿地松、火炬松离体培养植株再生技术的研究[D]. 南京: 南京林业大学, 2008.
[7]	姜福星, 魏丕伟, 寇亚平, 等. 探析植物体胚发生的关键基因[J]. 分子植物育种, 2017, 15(4): 1304-1311.
[8]	QUIROZ-FIGUEROA F R, ROJAS-HERRERA R, GALAZ-AVALOS R M, et al. Embryo production through somatic embryogenesis can be used to study cell differentiation in plants[J]. Plant Cell Tissue Organ Cult, 2006, 86(3): 285-301. doi: 10.1007/s11240-006-9139-6
[9]	KIM Y W, MOON H K. Enhancement of somatic embryogenesis and plant regeneration in Japanese red pine (Pinus densiflora)[J]. Plant Biotechnol Rep, 2014, 8(3): 259-266. doi: 10.1007/s11816-014-0319-2
[10]	PULLMAN G S, BUCALO K. Plant embryo culture[M]. New York: Humana Press, 2011: 267-291.
[11]	PULLMAN G S, MEIN J, JOHNSON S, et al. Gibberellin inhibitors improve embryogenic tissue initiation in conifers[J]. Plant Cell Rep, 2005, 23(9): 596-605. doi: 10.1007/s00299-004-0880-1
[12]	BONGA J M, PARK Y S, OVERTON C, et al. Optimized somatic embryogenesis in Pinus strobus L.[J]. In Vitro Cell Dev Biol Plant, 2013, 37(3): 392-399.
[13]	BECWAR M, CLARK J, SWINTON L S, et al. Liquid-based method for producing plant embryos: US 8313946B2[P]. 2012-11-20.
[14]	GUPTA P K, HOLMSTROM D. Protocol for somatic embryogenesis in woody plants[M]. Dordrecht: Springer, 2005: 573-575.
[15]	李清清, 叶建仁, 吴小芹. 黑松合子胚和体细胞胚发育阶段的形态特征比较[J]. 林业科技开发, 2012, 26(5): 21-23. doi: 10.3969/j.issn.1000-8101.2012.05.005
[16]	LIAO Y K, AMERSON H V. Slash pine (Pinus elliottii Engelm.) somatic embryogenesis: I: Initiation of embryogenic cultures from immature zygotic embryos[J]. New Forest, 1995, 10(2): 145-163.
[17]	吴涛, 陈少瑜, 陈芳, 等. 云南松合子胚发育及形态特征[J]. 西北林学院学报, 2008, 23(6): 91-93.
[18]	吕守芳, 张守攻, 齐力旺, 等. 日本落叶松体细胞胚胎发生的研究[J]. 林业科学, 2005, 41(2): 48-52. doi: 10.3321/j.issn:1001-7488.2005.02.008
[19]	张建伟, 王军辉, 李青粉, 等. 云杉未成熟合子胚诱导体细胞胚胎发生[J]. 林业科学, 2014, 50(4): 39-46.
[20]	RAMAROSANDRATANA A V, STADEN J V. Tissue position, explant orientation and naphtha aleneacetic acid (NAA) affect initiation of somatic embryos and callus proliferation in Norway spruce (Picea abies)[J]. Plant Cell Tissue Organ Cult, 2003, 74(3): 249-255. doi: 10.1023/A:1024022226381
[21]	丛建民, 沈海龙, 李玉花, 等. 水曲柳体胚发生过程中不同状态类型外植体的生理生化状态[J]. 华南农业大学学报, 2012, 33(1): 48-52. doi: 10.3969/j.issn.1001-411X.2012.01.010
[22]	徐春香, PANIS B, STROSSE H, 等. 体胚发生途径‘大矮蕉’植株的再生[J]. 华南农业大学学报, 2004, 25(2): 63-66. doi: 10.3969/j.issn.1001-411X.2004.02.017
[23]	徐春香, PANIS B, STROSSE H, 等. 香蕉胚性愈伤组织的诱导及胚性细胞悬浮系的建立[J]. 华南农业大学学报, 2004, 25(1): 70-73. doi: 10.3969/j.issn.1001-411X.2004.01.019
[24]	PULLMAN G S, NAMJOSHI K, ZHANG Y. Somatic embryogenesis in loblolly pine (Pinus taeda L.): Improving culture initiation with abscisic acid and silver nitrate[J]. Plant Cell Rep, 2003, 22(2): 85-95. doi: 10.1007/s00299-003-0673-y
[25]	PULLMAN G S, BUCALO K. Pine somatic embryogenesis: analyses of seed tissue and medium to improve protocol development[J]. New Forest, 2014, 45(3): 353-377. doi: 10.1007/s11056-014-9407-y
[26]	GUPTA P K, DURZAN D J. Shoot multiplication from mature trees of douglas-fir (Pseudotsuga menziesii) and sugar pine (Pinus lambertiana)[J]. Plant Cell Rep, 1985, 4(4): 177-179. doi: 10.1007/BF00269282
[27]	LITVAY J D, VERMA D C, JOHNSON M A. Influence of a loblolly pine (Pinus taeda L.) culture medium and its components on growth and somatic embryogenesis of the wild carrot (Daucus carota L.)[J]. Plant Cell Rep, 1985, 4(6): 325-328. doi: 10.1007/BF00269890
[28]	NUNES S, MARUM L, FARINHA N, et al. Somatic embryogenesis of hybrid Pinus elliottii var. elliottii×P. caribaea var. hondurensis and ploidy assessment of somatic plants[J]. Plant Cell Tissue Organ Cult, 2018, 132(1): 71-84. doi: 10.1007/s11240-017-1311-7
[29]	陈金慧, 施季森, 诸葛强, 等. 植物体细胞胚胎发生机理的研究进展[J]. 南京林业大学学报(自然科学版), 2003, 27(1): 75-80. doi: 10.3969/j.issn.1000-2006.2003.01.018
[30]	杨玲. 花楸种子生物学和体细胞胚发生体系研究[D]. 哈尔滨: 东北林业大学, 2007.
[31]	王高. 红松体细胞胚胎发生及超低温保存技术研究[D]. 上海: 上海交通大学, 2009.
[32]	NEWTON R J, TANG W, JAIN S M. Slash pine (Pinus elliottii Engelm.)[M]. Berlin Heidelberg: Springer, 2005: 1-10.
[33]	JAIN S M, DONG N, NEWTON R J. Somatic embryogenesis in slash pine (Pinus elliottii) from immature embryos cultured in vitro[J]. Plant Sci, 1989, 65(2): 233-241. doi: 10.1016/0168-9452(89)90070-8
[34]	唐巍, 欧阳藩, 郭仲琛. 湿地松体细胞胚胎发生和植株再生[J]. 植物资源与环境学报, 1997, 6(2): 8-11.

Cited By

Get Citation

PDF

XML

Article views (1700) PDF downloads (1549)

Turn off MathJax

Article Contents

Abstract

References

Somatic embryogenesis and plantlet regeneration in Pinus elliottiiand its hybrids

Abstract

References

Catalog

Export File

Citation

Format

Content