The factors influencing the embryogenic callus initiation and somatic embryo yield in Pinus elliottii resistant to pine needle brown spot disease

doi:10.12302/j.issn.1000-2006.202312024

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2025, Vol. 49 ›› Issue (1): 87-94.doi: 10.12302/j.issn.1000-2006.202312024

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The factors influencing the embryogenic callus initiation and somatic embryo yield in Pinus elliottii resistant to pine needle brown spot disease

KE Xin, FEI Qi, XIA Xinrui, YE Jianren, ZHU Lihua^*()

Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry and Grassland, Nanjing Forestry University, Nanjing 210037, China

Received:2023-12-25 Revised:2024-02-07 Online:2025-01-30 Published:2025-01-21
Contact: ZHU Lihua E-mail:lhzhu@njfu.com.cn

Abstract

Abstract:

【Objective】The somatic embryogenesis induction system of resistant Pinus elliottii is established and optimized to provide technical support for the large-scale rapid propagation of high-resistant materials of P. elliottii. 【Method】 The immature zygotic embryos from four families of P. elliottii are used to explore the effects of different families on the embryogenic callus initiation rate. The embryogenic callus with good proliferation rate and generating large quantity of embryo suspensor mass is selected to test somatic embryo maturation. The maturation conditions of somatic embryos are optimized based on genotype, phytosulfokine (PSK) concentration, and subculture times. 【Result】A significant difference is observed in the initiation frequencies of embryogenic callus among the four families (P < 0.05). Family No. 30 has the highest initiation rate (29.8 ± 5.6)%, while family No. 32 has the lowest (7.3 ± 3.4)%. A total of 37 embryogenic cell lines with stable proliferation are obtained. Of these, 25.7% of 35 cell lines produce matured somatic embryos, and the yields (number of somatic embryos) in 2007-3 and 2007-5 are significantly higher than that of other cell lines, with an average yield of (91.3 ± 7.0) and (89.8 ± 8.4) cotyledonary somatic embryos per mL, respectively, while the remaining 74.3% does not produce any somatic embryos. The addition of PSK significantly affects the yield of somatic embryos of resistant P. elliottii, and embryo production is considerably increased when the PSK concentration is 1 mg/L. The number of mature somatic embryos decreases with the increase in subcultures. The number of mature somatic embryos of cell lines 2007-3 gradually decreases after eight subcultures but maintains embryogenic competence in the 20^th subculture. For cell lines 2007-5, the number of mature somatic embryos reduces as subcultures rise, ultimately losing maturation capacity after 20 subcultures. 【Conclusion】The initiation of embryogenic callus is greatly influenced by the genotype. Genotype and PSK concentration significantly affect the somatic embryo yield of resistant P. elliottii. The yield of somatic embryos decreases with the increase in subcultures. The somatic embryogenesis technology system of resistant P. elliottii is optimized to provide technical support for the somatic embryogenesis of excellent genotypes of P. elliottii.

Key words: Pinus elliottii, somatic embryogenesis, genotype, phytosulfokine, somatic embryo maturation, breeding for disease resistance

CLC Number:

Q943
S722

KE Xin, FEI Qi, XIA Xinrui, YE Jianren, ZHU Lihua. The factors influencing the embryogenic callus initiation and somatic embryo yield in Pinus elliottii resistant to pine needle brown spot disease[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2025, 49(1): 87-94.

Figures/Tables 5

Fig. 1

Table 1

Development stage and yield of somatic embryo of 37 embryogenic cell lines in resistant Pinus elliottii"

家系号 family No.	细胞系 cell line	显微结构 microstructure	成熟体胚数量/ (个·mL^-1) number of somatic embryos	体胚形态 the morphology of somatic embryos	家系号 family No.	细胞系 cell line	显微结构 microstructure	成熟体胚数量/ (个·mL^-1) number of somatic embryos	体胚形态 the morphology of somatic embryos
7	2007-1	PEM Ⅰ、Ⅱ	0	未产生体胚	27	2027-5	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-2	PEM Ⅱ、Ⅲ	0	未产生体胚		2027-6	PEM Ⅰ、Ⅱ	2.3±1.3 e	白色、胚轴膨大、子叶闭合
	2007-3	SE	91.3±7.0 a	白色、胚轴伸长、子叶展开		2027-7	PEM Ⅱ、Ⅲ	0	未产生体胚
	2007-4	PEM Ⅱ、Ⅲ	0	未产生体胚		2027-8	PEM Ⅱ、Ⅲ	0	未产生体胚
	2007-5	PEM Ⅰ、Ⅱ	89.8±8.4 a	白色、胚轴伸长、子叶展开		2027-9	PEM Ⅱ、Ⅲ	0	未产生体胚
	2007-8	PEM Ⅱ、Ⅲ	2.0±0.8 e	黄色、胚轴粗壮、子叶闭合		2027-10	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-9	PEM Ⅱ、Ⅲ	0	未产生体胚		2027-11	PEM Ⅰ、Ⅱ	8.0±2.5 d	黄色、胚轴粗壮、子叶闭合
	2007-10	SE	28.0±2.9 b	黄绿色、胚轴粗壮、子叶展开		2027-12	PEM Ⅱ、Ⅲ	9.0±1.6 d	白色、胚轴膨大、子叶展开
	2007-12	PEM Ⅱ、Ⅲ	0	未产生体胚	30	2030-2	PEM Ⅱ、Ⅲ	0	未产生体胚
	2007-14	PEM Ⅱ、Ⅲ	0	未产生体胚		2030-5	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-15	PEM Ⅰ、Ⅱ	0	未产生体胚		2030-7	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-17	SE	0	未产生体胚		2030-9	SE	0	未产生体胚
	2007-19	SE	0	未产生体胚		2030-11	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-21	PEM Ⅱ、Ⅲ	0	未产生体胚		2030-12	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-24	SE	21.3±2.6 c	黄绿色、胚轴膨大、子叶闭合		2030-13	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-26	PEM Ⅰ、Ⅱ	0	未产生体胚		2030-14	PEM Ⅰ、Ⅱ	0	未产生体胚
	2007-27	SE	20.7±3.1 c	黄色、胚轴伸长、子叶展开	32	2032-1	PEM Ⅰ、Ⅱ	—	—
27	2027-1	SE	0	未产生体胚		2032-2	PEM Ⅰ、Ⅱ	—	—
	2027-4	PEM Ⅱ、Ⅲ	0	未产生体胚

Table 1

Fig. 2

Fig. 3

Fig. 4

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The factors influencing the embryogenic callus initiation and somatic embryo yield in Pinus elliottii resistant to pine needle brown spot disease

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