Response Surface Optimization of Culture Conditions for the Formation of Early Embryos in Larix olgensis

doi:10.7525/j.issn.1673-5102.2017.04.018

Abstract

Abstract: With the larch embryogenic callus, through the Design Box-Behnken design test, we established a response surface model based on the somatic embryogenesis. The culture conditions of somatic embryo early maturation was optimized in Larix olgensis. The relative ion concentration(P=0.007), the inositol concentration(P=0.000), the culture period(P=0.000) and the interaction between inositol content and culture period(P=0.007) effect on the number of somatic embryos of L.olgensis were extremely significant. Cultivation of Larix somatic embryo maturation was 337.04 ind·g^-1 at the early stage of the most optimum conditions for relative ion content of 26.767 7% BM basic culture medium, inositol 10.454 5 g·L^-1, culture time 12.656 6 d, and the number of somatic embryos average. In order to facilitate the practical operation, the culture condition was changed to the relative ion concentration of 25% BM, inositol 10 g·L^-1, incubation period of 13 d, then the average number of somatic embryos was 336.46 ind·g^-1, that was consistent with the predicted value 336.29 ind·g^-1. Compared with previous test, the quantity and quality of L.olgensis somatic embryos were increased significantly.

Key words: Larix, Larix olgensis, response surface optimization, somatic embryogenesis, early embryo formation

CLC Number:

SONG Yue, LI Shu-Juan, BAI Xiao-Ming, ZHANG Han-Guo. Response Surface Optimization of Culture Conditions for the Formation of Early Embryos in Larix olgensis[J]. Bulletin of Botanical Research, 2017, 37(4): 613-620.

References

1. 王伟达,李成浩,杨静莉,等. 杂种落叶松未成熟胚的体细胞胚发生和植株再生[J]. 林业科学,2009,45(8):34-38.Wang W D,Li C H,Yang J L,et al. Somatic embryogenesis and plantlet regeneration from immature zygotic embryos of hybrid larch[J]. Scientia Silvae Sinicae,2009,45(8):34-38.
2. 杨书文. 落叶松的遗传改良[M]. 哈尔滨:东北林业大学出版社,1994.Yang S W. Genetic improvement of larch[M]. Harbin:Northeast Forestry University Press,1994.
3. 刘足根,朱教君,袁小兰,等. 辽东山区长白落叶松(Larix olgensis)种子雨和种子库[J]. 生态学报,2007,27(2):579-587.Liu Z G,Zhu J J,Yuan X L,et al. On seed rain and soil seed bank of Larix olgensis in montane regions of eastern Liaoning Province,China[J]. Acta Ecologica Sinica,2007,27(2):579-587.
4. 宋跃,甄诚,张含国,等. 长白落叶松胚性愈伤组织诱导及体细胞胚胎发生[J]. 林业科学,2016,52(10):45-54.Song Y,Zhen C,Zhang H G,et al. Embryogenic callus induction and somatic embryogenesis from immature zygotic embryos of Larix olgensis[J]. Scientia Silvae Sinicae,2016,52(10):45-54.
5. Von Aderkas P,Klimaszewska K,Bonga J M. Diploid and haploid embryogenesis in Larix leptolepis,L. decidua,and their reciprocal hybrids[J]. Canadian Journal of Forest Research,1990,20(1):9-14.
6. 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.Lü S F,Zhang S G,Qi L W,et al. Somatic embryogenesis from immature embryos of Larix kaempferi[J]. Scientia Silvae Sinicae,2005,41(2):48-52.
7. 汪小雄,杨映根. 日本落叶松体细胞胚胎发生的研究[J]. 安徽农业科学,2010,38(4):2118-2121.Wang X X,Yang Y G. Study on the somatic embryogenesis of Larix leptolepis[J]. Journal of Anhui Agricultural Sciences,2010,38(4):2118-2121.
8. 齐力旺. 华北落叶松体细胞胚胎发生与遗传转化系统建立的研究[D]. 北京:中国林业科学研究院,2000.Qi L W. Studies on the somatic embryogenesis and establishment of experimental system in Larix principis-Rupprechtii[D]. Beijing:Chinese Academy of Forestry,2000.
9. Klimaszewska K. Plantlet development from immature zygotic embryos of hybrid larch through somatic embryogenesis[J]. Plant Science,1989,63(1):95-103.
10. Lelu M A,Bastien C,Klimaszewska K,et al. An improved method for somatic plantlet production in hybrid larch(Larix×leptoeuropaea):Part 1. Somatic embryo maturation[J]. Plant Cell,Tissue and Organ Culture,1994,36(1):107-115.
11. 王伟达,李成浩,杨静莉,等. 不同植物生长调节物质对杂种落叶松胚性愈伤组织增殖的影响[J]. 东北林业大学学报,2008,36(9):5-7.Wang W D,Li C H,Yang J L,et al. Effect of different plant growth regulators on embryogenic callus proliferation of hybrid larix[J]. Journal of Northeast Forestry University,2008,36(9):5-7.
12. 中国林业科学研究院林业研究所. 一种提高针叶树种体细胞胚胎发生数量与质量的培养基:中国,CN1733906A[P]. 2006-02-15.INST Forestry CAF. Medium for improving quantity and quality of conifer somatic embryogenesis:Chinese,CN1733906A[P]. 2006-02-15.
13. 中国林业科学研究院林业研究所. 一种改良针叶树种胚性愈伤组织质量的培养基:中国,CN1733905A[P]. 2006-02-15.INST Forestry CAF. Culture medium for improving conifer embryogenic callus quality:Chinese,CN1733905A[P]. 2006-02-15.
14. 罗猛,宋卓悦,胡娇阳,等. 超声法提取山里红叶总黄酮及其抗氧化活性研究[J]. 植物研究,2015,35(4):632-637.Luo M,Song Z Y,Hu J Y,et al. Optimization of ultrasonic-assisted extraction of total flavonoids from Crataegus pinnatifida leaves and its antioxidant activities[J]. Bulletin of Botanical Research,2015,35(4):632-637.
15. 郝学财,余晓斌,刘志钰,等. 响应面方法在优化微生物培养基中的应用[J]. 食品研究与开发,2006,27(1):38-41.Hao X C,Yu X B,Liu Z Y,et al. The application of response surface methodology in optimization of microbial media[J]. Food Research and Development,2006,27(1):38-41.
16. 东北林业大学. 一种提高长白落叶松体细胞胚发生和植株再生效率的过渡培养基及培养方法:中国,CN105409767A[P]. 2016-03-23.Northeast Forestry University. Transitional culture medium improving somatic embryogenesis and plant regeneration efficiency of Larix olgensis and culture method:Chinese,CN105409767A[P]. 2016-03-23.
17. 李淑娟,詹亚光,杨传平,等. 基于响应面法的白蜡属花粉离体萌发培养基优化[J]. 植物学报,2009,44(2):223-229.Li S J,Zhan Y G,Yang C P,et al. Response surface methodology to determine the optimal medium for pollen germination for ash[J]. Chinese Bulletin of Botany,2009,44(2):223-229.
18. Amenaghawon N A,Nwaru K I,Aisien F A,et al. Application of box-behnken design for the optimization of citric acid production from corn starch using Aspergillus niger[J]. British Biotechnology Journal,2013,3(3):236-245.
19. 王化,周丽萍,李梦莎,等. 微波辅助提取暴马丁香中紫丁香苷和橄榄苦苷的工艺研究[J]. 植物研究,2016,36(1):141-145.Wang H,Zhou L P,Li M S,et al. Optimization of microwave-assisted extraction of syringin and oleuropein from Syringa reticulata[J]. Bulletin of Botanical Research,2016,36(1):141-145.
20. 刘燕,陈小银,杨丽丽,等. 响应面优化菊芋菊糖的提取工艺研究[J]. 植物研究,2016,36(4):627-633.Liu Y,Chen X Y,Yang L L,et al. Extraction process optimization of inulin from Helianthus tuberosus L. by response surface methodology[J]. Bulletin of Botanical Research,2016,36(4):627-633.
21. 林建原,季丽红. 响应面优化银杏叶中黄酮的提取工艺[J]. 中国食品学报,2013,13(2):83-90.Lin J Y,Ji L H. Optimization of flavonoids from Ginkgo biloba using respone surface analysis[J]. Journal of Chinese Institute of Food Science and Technology,2013,13(2):83-90.
22. 崔凯荣,戴若兰. 植物体细胞胚发生的分子生物学[M]. 北京:科学出版社,2000.Cui K R,Dai R L. Molecular biology of plant somatic embryogenesis[M]. Beijing:Science Press,2000.
23. 汪小雄,卢龙斗,郝怀庆,等. 松杉类植物体细胞胚发育机理的研究进展[J]. 西北植物学报,2006,26(9):1965-1972.Wang X X,Lu L D,Hao H Q,et al. Research advances about developmental mechanism of somatic embryos in conifers[J]. Acta Botanica Boreali-Occidentalia Sinica,2006,26(9):1965-1972.
24. Singh H. Embryology of gymnosperms[M]. Berlin:Borntrager,1978.
25. Filonova L H,Bozhkov P V,Von Arnold S. Developmental pathway of somatic embryogenesis in Picea abies as revealed by time-lapse tracking[J]. Journal of Experimental Botany,2000,51(343):249-264.
26. Filonova L H,Bozhkov P V,Brukhin V B,et al. Two waves of programmed cell death occur during formation and development of somatic embryos in the gymnosperm,Norway spruce[J]. Journal of Cell Science,2000,113(24):4399-4411.
27. 向太和,钟华鑫,梁海曼,等. 水稻胚性悬浮细胞系建立过程中的生理生化变化[J]. 作物学报,1995,21(2):223-119.Xiang T H,Zhong H X,Liang H M,et al. Physiological and biochemical changes during the establishment of embryogenic suspension cell lines of rice(Oryza sativa L.)[J]. Acta Agronomica Sinica,1995,21(2):223-119.
28. Irvine R F,Schell M J. Back in the water:the return of the inositol phosphates[J]. Nature Reviews Molecular Cell Biology,2001,2(5):327-338.
29. Hall P J,Bandurski R S. [³H]Indole-3-acetyl-myo-inositol hydrolysis by extracts of Zea mays L. vegetative tissue[J]. Plant Physiology,1986,80(2):374-377.
30. Peskan T,Westermann M,Oelmüller R. Identification of low-density Triton X-100-insoluble plasma membrane microdomains in higher plants[J]. European Journal of Biochemistry,2000,267(24):6989-6995.
31. Tiede A,Bastisch I,Schubert J P,et al. Biosynthesis of glycosylphosphatidylinositols in mammals and unicellular microbes[J]. Biological Chemistry,1999,380(5):503-523.
32. Borner G H H,Sherrier D J,Weimar T,et al. Analysis of detergent-resistant membranes in Arabidopsis. Evidence for plasma membrane lipid rafts[J]. Plant Physiology,2005,137(1):104-116.
33. Luo Y,Qin G,Zhang J,et al. D-myo-inositol-3-phosphate affects phosphatidylinositol-mediated endomembrane function in Arabidopsis and is essential for auxin-regulated embryogenesis[J]. The Plant Cell,2011,23(4):1352-1372.
34. 王静. 利用胚性愈伤组织进行落叶松遗传转化体系的研究[D]. 北京:中国林业科学研究院,2007.Wang J. Studies on the transformation of Larix based on the system of somatic embryogenesis[D]. Beijing:Chinese Academy of Forestry,2007.

[1]	Yuhui REN, Shuai NIE, Chunxue PENG, Ling YANG, Hailong SHEN. Effects of Hormone Combinations, Carbon Source Type and Proliferation Cycle on Embryogenic Callus Proliferation of Pinus koraiensis [J]. Bulletin of Botanical Research, 2022, 42(4): 704-712.
[2]	Xueyan ZHOU, Biying WANG, Xuefeng HAO, Xinguo HU, Jiangtao WU, Kai LANG, Qinbo HU, Xiyang ZHAO. Genetic Variation and Joint Selection of Growth and Wood Traits in Half-sib Families of Larix olgensis [J]. Bulletin of Botanical Research, 2022, 42(3): 383-393.
[3]	Tongtong Sun, Zhongjuan Wang, Xiujuan Zhang, Shihan Yang, Xiaoqiang Chen, Ying Zhang. Optimization on Extraction of Total Phenols by Response Surface from Saxifraga stolonifera and Its Correlation in Inhibiting the Activity of PLA₂ in Deinagkistrodon acutus Venom [J]. Bulletin of Botanical Research, 2022, 42(2): 259-267.
[4]	Yuning Yang, Hao Dong, Shiwei Dong, Nairui Wang, Yue Song, Hanguo Zhang, Shujuan Li. Cloning and Expression Analysis of Transcription Factor LobHLH34 from Larix olgensis [J]. Bulletin of Botanical Research, 2022, 42(1): 112-120.
[5]	Dandan Zhang, Xiang Li, Biying Wang, Xihe Wang, Quan Sun, Yunyang Wu, Pingyang Li, Deyao Li, Yulei Li, Xiyang Zhao. Variation Analysis of Growth Traits of Larix olgensis Parental Clones in Seed Orchards [J]. Bulletin of Botanical Research, 2022, 42(1): 130-137.
[6]	Chun-Xue PENG, Xue-Mei CUI, Hai-Long SHEN. Callus Induction and Somatic Embryogenesis and Physiological State Analysis from Mature Zygotic Embryo Explant of Syringa reticulata var. mandshurica [J]. Bulletin of Botanical Research, 2021, 41(4): 557-563.
[7]	Fang WANG, Lu-Ping JIANG, Qin-Hui ZHANG, Zhi-Min LU, Yu-Chun YANG, Jian-Qiu ZHANG, Xi-Yang ZHAO. Stability Analysis and Genetic Variation in Seedling Growth of 51 Larix olgensis Clones from Different Sites [J]. Bulletin of Botanical Research, 2021, 41(3): 336-343.
[8]	Ting-Yan LIU, Long-Fei HAO, Xu-Fu WANG, Hai-Xia YAN, Shu-Lan BAI. Effects of Nitrogen Deposition and Ectomycorrhizal Fungi on Root Architecture and Rhizosphere Soil Enzyme Activities of Larix olgensis Seedlings [J]. Bulletin of Botanical Research, 2021, 41(1): 145-151.
[9]	ZHANG Lei, XIONG Huan-Huan, CAO Qing, ZHAO Jia-Li, ZHANG Han-Guo. Drought Resistance of Larch NAC Gene by Transient Genetic Transformation [J]. Bulletin of Botanical Research, 2020, 40(3): 394-400.
[10]	BAI Xiao-Ming, DONG Shi-Wei, YANG Yu-Ning, SONG Yue, ZHANG Han-Guo, LI Shu-Juan. Cloning and Expression Analysis of Catalase Gene(LoCAT1) from Larix olgensis [J]. Bulletin of Botanical Research, 2019, 39(4): 539-546.
[11]	PENG Chun-Xue, CUI Xue-Mei, LIU Chun-Ping, YANG Ling, WANG Qiu-Shui, SHEN Hai-Long. Physiological Analysis of Three Anti-Browning Agents Affecting Somatic Embryogenesis in Fraxinus mandshurica [J]. Bulletin of Botanical Research, 2019, 39(2): 252-258.
[12]	YANG Ling, LIU Hong-Nan, ZHANG Dong-Yan, WEI Cheng, SHEN Hai-Long. Effect of Plant Growth Regulators and Osmoticums on Somatic Embryogenesis of Fraxinus mandshurica Rupr. [J]. Bulletin of Botanical Research, 2017, 37(5): 682-689.
[13]	HUANG Zhen1,2；LI Yun1,2；LI Yuan1,2；WANG Pei-Qi1,2；KANG Xiang-Yang1,2. Establishment and Regeneration of Suspension Cell Lines in Hybirds of （Populus pseudo-simonii×P.nigra* ‘Zhenyin3#’）×P.×beijingensis [J]. Bulletin of Botanical Research, 2015, 35(5): 696-703.
[14]	LIAO Jing;FANG Yan-Ming;YU Mu-Kui. Preliminary Report on Somatic Embryogenesis from Stem of Quercus acutissima* Carr. and Histological Observation [J]. Bulletin of Botanical Research, 2011, 31(5): 575-578.
[15]	YANG Ling;SHEN Hai-Long;LIU Chun-Ping;ZHAI Xiao-Jie. Somatic Embryogenesis from Mature Zygotic Embryo Explants of Sorbus pohuashanensis* Hedl [J]. Bulletin of Botanical Research, 2010, 30(2): 174-179.