Populus alba xP .berolinensis hormone autotrophic cell line capable of realizing plant regeneration
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A cell line, autotrophic technology that can be used in the biological field to solve problems such as non-redifferentiation
Active Publication Date: 2021-02-12
NORTHEAST FORESTRY UNIVERSITY
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The tobacco BY2 cell line in plants, since its discovery in the 1970s (kato 1972), has played an important role in revealing the cell cycle (delporte et al. 2014; Harashima et al. 2007; Yu et al. 2006), cell metabolism (Issawi et al. 2017; Krystofova et al. 2012; Han et al. 2014), abiotic stress studies (Banu et al. 2009; Gai et al. 2011) and verification of gene function (Toussaint et al. 2017; Niczyj et al. 2016; Staub 2014 years; Han et al. 2014) and other mechanisms have also played an important role, and its disadvantage is that it cannot be redifferentiated to achieve plant regeneration
A variety of good plant suspension cell lines have been obtained, such as Mediterranean thistle (Cnicus benedictus L.) (Lattanzio et al. 2018) cell line, switchgrass (Panicum Virgatum) cell line (Rao et al. 2017), basmati rice (Pusa Basmati.Biotech) cell line (Sathish et al. 2018), among others, cell lines with good properties on forest trees have not been reported
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example 1
[0013] The acquisition of the Yang hormone autotrophic cell line of example 1 silver
[0014] 1) Pollen picking and callus induction of Populus yinzhong
[0015] In mid-April 2015, the pollen development period of Populus silvery in the campus of Northeast Forestry University was observed by DAIP staining. After the pollen in the mononuclear marginal stage was sterilized in the ultra-clean workbench, it was scattered and inoculated on a medium supplemented with 2mg / L 2, 4-D, 0.5mg / L KT, 30g / L sucrose and 7.5g / L agar In MS medium, callus was induced by culturing in the dark for 30 days.
[0016] 2) Subculture of callus and acquisition of hormone autotrophic cell lines
[0017] The obtained callus was subcultured once every 2 weeks, and after 3 consecutive subcultures, the callus with faster growth and better state was selected, and a loose light yellow callus with vigorous growth was obtained after 5 consecutive subcultures. Then it was transferred to MS medium containing on...
example 2
[0018] Suspension culture of example 2 hormone autotrophic cell line
[0019] The hormone autotrophic cell line with a fresh weight of 0.9-1.2g was placed in a 150mL glass Erlenmeyer flask containing only 50g / L sucrose 50mL MS liquid medium for culture, and the growth of the cell line within 18 days was counted, as shown in figure 2 d. It can be seen from the figure that the growth amount reached 0.45g on the 18th day, an increase of about 9.2 times, and continued to grow until it became viscous and filled the entire culture bottle. The cell line is subcultured every 15-20 days in the medium without any exogenous hormones, which can ensure the best growth state.
example 3
[0020] Plant regeneration of example 3 hormone autotrophic cell line
[0021] The subcultured hormonal autotrophic cell lines were inoculated on MS medium containing different plant growth regulators (Table 1) to induce bud differentiation. Adventitious buds grew in the medium of mg / L TDZ, but no adventitious buds appeared in the other medium settings, and some even browned and died. The induced adventitious buds were cultured for rooting and seedlings were obtained.
[0022] Table 1 Culture medium for inducing bud differentiation of cell lines
[0023]
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Abstract
The invention belongs to the technical field of plant biology, and mainly provides a populus alba xP .berolinensis hormone autotrophic cell line capable of realizing plant regeneration. A suspension culture system and an adventitious bud differentiation and rooting plant regeneration system are established; callus is induced from populus alba xP .berolinensis pollen in a mononuclear edging stage in an MS culture medium added with 2 mg/L of 2,4-D, 0.5 mg/L of KT, 30g/L of sucrose and 7.5 g/L of agar, and continuous subculture is carried out for 5 times to obtain light yellow callus with loose texture and vigorous growth; the callus still can rapidly grow in an MS culture medium only containing 30 g/L of sucrose and is the hormone autotrophic cell line; after the hormone autotrophic cell line is inoculated into a 150 mL glass triangular flask containing 50 mL of a MS liquid culture medium with 50 g/L of sucrose for suspension culture for 18 days, the growth amount is increased by about 9.2 times; culturing is carried out in an MS culture medium added with 0.05 mg/L of TDZ, 25g/L of sucrose and 7.5 g/L of agar for 3 months, and inducing is carried out to form a large number of adventitious buds; and the adventitious buds are inoculated into a rooting culture medium to be cultured for 30 days, rooting of the adventitious buds is achieved, and regenerated plants are formed.
Description
technical field [0001] The invention belongs to the field of biotechnology, and mainly relates to obtaining a hormone autotrophic cell line of poplar (Populus alba×P.berolinensis) by using a plant tissue culture method, the cell line is pale yellow, loose in texture, and has no plant growth regulation It can grow rapidly with the addition of the agent, and transfer to the differentiation medium can induce budding and realize plant regeneration. The acquisition of this cell line will provide a good tool for forest tree genetics and breeding, revealing forest tree growth and development, and stress mechanisms. Background technique [0002] Suspension cell lines with good homogeneity and rapid growth are ideal research tools in the fields of cell biology, genetics, and biological metabolism (Moscatiello et al. 2013). When it comes to some research content with complex biological processes and long experimental cycles, it is particularly important to choose appropriate research...
Claims
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