This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the ge...This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the genetic regeneration of maize and the verification of the genetic functions of maize.The factor experiment and orthogonal experiments were used to investigate the impacts of different genotypes,immature embryo size,shield orientation,2,4-D concentration,proline concentration,and folic acid concentration on the induction rate of embryogenic callus tissue.A sensitivity experiment testing glyphosate(Bar)and an antibiotic(Cefotaxime sodium)were also conducted.The results indicate that the immature embryos of inbred maize line GSH9901 were the most effective for callus tissue induction,and the immature embryos with a length of 1.6-2.0 mm produce the best result.The upward shield face is more successful for the formation of induced callus.Using orthogonal analysis,we found that the optimal combination for the induction system was A_(3)(2,4-D concentration 0.25 mg mL^(-1)),B_(1)C_(3)(proline concentration 0.8 mg mL^(-1)),and D 2(folate Concentration 0.5 mg mL^(-1))and the induction rate reached 84%.We found that cold storage at 4℃ for 1 d is more conducive for the formation of embryogenic callus than the other treatments tested.The sensitivity experiment for callus tissue screening revealed the critical concentration of glyphosate to be 10 mg ml^(-1),and the critical concentration of antibiotic is 250 mg ml^(-1).Using this combination of glyphosate and antibiotic resulted in regenerated plants.This study established the optimal conditions for immature embryo callus tissue induction in maize.展开更多
The embryos of maize(Zea mays L.)inbred lines GS02,GS07,GS08,GS11 and GS15 were used as receptor materials to optimize the receptor system from the aspects of genotype,medium components and stress(PEG6000,mannitol,sal...The embryos of maize(Zea mays L.)inbred lines GS02,GS07,GS08,GS11 and GS15 were used as receptor materials to optimize the receptor system from the aspects of genotype,medium components and stress(PEG6000,mannitol,salt and low phosphorus).The results showed that GS07 had the highest induction rate(95.2%).Orthogonal test analysis showed that the best combination of medium components in induction was A2B3C1D3(2),namely,the concentration of 2,4-dichlorophenoxy acetic acid(2,4-D)was 4 mg·mL^(-1),the concentration of L-Proline(L-Pro)was 0.8 mg·mL^(-1),and the concentration of silver nitrate(AgNO3)was 10 mg·mL^(-1)(or 5 mg·mL).Interestingly,we found that the optimal medium supplemented with 30 g·L^(-1)PEG6000 or 80 g·L^(-1)mannitol was suitable for antioxidant enzyme activity and malondialdehyde(MDA)content in GS07 callus.Exogenous 10 mmol·L^(-1)Ca^(2+)in the medium components with 100 mmol·L^(-1)sodium chloride(NaCl)could improve the activity of antioxidant enzymes in GS07 callus.Callus of GS07 could divide normally and grow well in medium components with 27 mg·L^(-1)KH_(2)PO_(4).This study enhanced the adaptability of maize callus to stress and optimized the culture conditions.展开更多
The induction,subculture and differentiation of callus from immature embryos of maize( Zea mays L.) inbred line were studied and optimized. The results revealed that,2 mg/L 2,4-D was the optimal concentration to induc...The induction,subculture and differentiation of callus from immature embryos of maize( Zea mays L.) inbred line were studied and optimized. The results revealed that,2 mg/L 2,4-D was the optimal concentration to induce embryonic callus. Calli induced from inbred Qi319 and LY92 had good morphology and high regeneration. The embryos of the two inbred lines were selected as explants to establish efficient and stable tissue culture and transformation system.展开更多
Betula platyphylla is a native tree species in northern China that has high economic and medicinal value.We developed an efficient protocol for the induction of somatic embryogenesis in B.platyphalla from immature zyg...Betula platyphylla is a native tree species in northern China that has high economic and medicinal value.We developed an efficient protocol for the induction of somatic embryogenesis in B.platyphalla from immature zygotic embryos and assessed the effects of explant type,genotype,and plant growth regulators(PGRs)on embryogenic callus induction.Among the various explants evaluated,embryogenic callus was only produced from mature and immature zygotic embryos on medium with added 2,4-dichlorophenoxyacetic acid(2,4-D).Supplementation of 2,4-D-containing medium with cytokinins increased the frequency of embryogenic callus induction.On the 20 days after pollination,immature zygotic embryos that had been collected in mid-May yielded embryogenic tissue at the highest frequency(16.8%)when cultured on half-strength MS medium supplemented with 2.0 mg L^(-1)2,4-D and 0.2 mg L^(-1)6-benzylaminopurine(6-BA).The process of proliferation of embryogenic callus,somatic embryo formation,and subsequent plantlet conversion occurred under optimal culture conditions.When regenerated plants weretransplanted to soil,95%of them developed normally and grew vigorously.This somatic embryogenesis system required 3–4 months for the regeneration of B.platyphalla plantlets from immature zygotic embryos.展开更多
Immature embryos of inbred maize(Zea mays)lines(H8183,H8184,and H8185)were used for Agrobacterium infection.We used theβ-glucuronidase gene(GUS)as the target gene and the glufosinate resistance gene(bar)as the select...Immature embryos of inbred maize(Zea mays)lines(H8183,H8184,and H8185)were used for Agrobacterium infection.We used theβ-glucuronidase gene(GUS)as the target gene and the glufosinate resistance gene(bar)as the selection marker.We conducted research on several aspects,such as different genotypes,coculture conditions,screening agent concentrations,and concentrations of indole-3-butytric acid(IBA),6-benzylaminopurine(6-BA),and ascorbic acid(Vc)in the differentiation medium.We optimized the genetic transformation system,and the obtained results indicated that among the three lines studied,the induction rate of H8185 was the highest at 93.2%,followed by H8184,with H8183 having the lowest induction rate(80.1%).The best coculture method was that using the N6 coculture medium layered with a sterile filter paper.Using orthogonal analysis,we found that the optimal combination of the three factors in the differentiation medium was A_(3)(1 mg mL^(−1)IBA),B_(3)C_(1)(1.6 mg mL^(−1)6-BA),and D_(3)(1.5 mg mL^(−1)Vc).Through GUS staining analysis,Bar test-strip analysis,and polymerase chain reaction,five transgenic plants were finally obtained.This study established the optimal conditions for genetic transformation in maize.展开更多
基金This research was funded by Jilin province science and technology research projects(20170204005NY)Jilin Science and Technology Development Plan Major Science and Technology R&D Project(20180201029NY)+2 种基金Jilin Province Science and Technology Development Plan Project(20190802012ZG)Jilin Province Natural Science Foundation(20190201168JC)a thirteenth five-year plan for the Education Department of Jilin Province(JJKH20180661KJ)were jointly funded.
文摘This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the genetic regeneration of maize and the verification of the genetic functions of maize.The factor experiment and orthogonal experiments were used to investigate the impacts of different genotypes,immature embryo size,shield orientation,2,4-D concentration,proline concentration,and folic acid concentration on the induction rate of embryogenic callus tissue.A sensitivity experiment testing glyphosate(Bar)and an antibiotic(Cefotaxime sodium)were also conducted.The results indicate that the immature embryos of inbred maize line GSH9901 were the most effective for callus tissue induction,and the immature embryos with a length of 1.6-2.0 mm produce the best result.The upward shield face is more successful for the formation of induced callus.Using orthogonal analysis,we found that the optimal combination for the induction system was A_(3)(2,4-D concentration 0.25 mg mL^(-1)),B_(1)C_(3)(proline concentration 0.8 mg mL^(-1)),and D 2(folate Concentration 0.5 mg mL^(-1))and the induction rate reached 84%.We found that cold storage at 4℃ for 1 d is more conducive for the formation of embryogenic callus than the other treatments tested.The sensitivity experiment for callus tissue screening revealed the critical concentration of glyphosate to be 10 mg ml^(-1),and the critical concentration of antibiotic is 250 mg ml^(-1).Using this combination of glyphosate and antibiotic resulted in regenerated plants.This study established the optimal conditions for immature embryo callus tissue induction in maize.
基金the Science and Technology Project of Jilin Provincial Department of Education[JJKH20210351KJ,JJKH20210346KJ]Jilin Province Science and Technology Development Plan Project[20200402023NC]。
文摘The embryos of maize(Zea mays L.)inbred lines GS02,GS07,GS08,GS11 and GS15 were used as receptor materials to optimize the receptor system from the aspects of genotype,medium components and stress(PEG6000,mannitol,salt and low phosphorus).The results showed that GS07 had the highest induction rate(95.2%).Orthogonal test analysis showed that the best combination of medium components in induction was A2B3C1D3(2),namely,the concentration of 2,4-dichlorophenoxy acetic acid(2,4-D)was 4 mg·mL^(-1),the concentration of L-Proline(L-Pro)was 0.8 mg·mL^(-1),and the concentration of silver nitrate(AgNO3)was 10 mg·mL^(-1)(or 5 mg·mL).Interestingly,we found that the optimal medium supplemented with 30 g·L^(-1)PEG6000 or 80 g·L^(-1)mannitol was suitable for antioxidant enzyme activity and malondialdehyde(MDA)content in GS07 callus.Exogenous 10 mmol·L^(-1)Ca^(2+)in the medium components with 100 mmol·L^(-1)sodium chloride(NaCl)could improve the activity of antioxidant enzymes in GS07 callus.Callus of GS07 could divide normally and grow well in medium components with 27 mg·L^(-1)KH_(2)PO_(4).This study enhanced the adaptability of maize callus to stress and optimized the culture conditions.
基金Supported by Science and Technology Development Plan of Shandong ProvinceNational Key Laboratory Open Project of Crop Biology(2014KF11)
文摘The induction,subculture and differentiation of callus from immature embryos of maize( Zea mays L.) inbred line were studied and optimized. The results revealed that,2 mg/L 2,4-D was the optimal concentration to induce embryonic callus. Calli induced from inbred Qi319 and LY92 had good morphology and high regeneration. The embryos of the two inbred lines were selected as explants to establish efficient and stable tissue culture and transformation system.
基金supported by the National Key Research and Development Programme of China(No.2017YFD0600603)the 111 Project(No.B16010)the Heilongjiang Touyan Innovation Team Program(Tree Genetics and Breeding Innovation Team)。
文摘Betula platyphylla is a native tree species in northern China that has high economic and medicinal value.We developed an efficient protocol for the induction of somatic embryogenesis in B.platyphalla from immature zygotic embryos and assessed the effects of explant type,genotype,and plant growth regulators(PGRs)on embryogenic callus induction.Among the various explants evaluated,embryogenic callus was only produced from mature and immature zygotic embryos on medium with added 2,4-dichlorophenoxyacetic acid(2,4-D).Supplementation of 2,4-D-containing medium with cytokinins increased the frequency of embryogenic callus induction.On the 20 days after pollination,immature zygotic embryos that had been collected in mid-May yielded embryogenic tissue at the highest frequency(16.8%)when cultured on half-strength MS medium supplemented with 2.0 mg L^(-1)2,4-D and 0.2 mg L^(-1)6-benzylaminopurine(6-BA).The process of proliferation of embryogenic callus,somatic embryo formation,and subsequent plantlet conversion occurred under optimal culture conditions.When regenerated plants weretransplanted to soil,95%of them developed normally and grew vigorously.This somatic embryogenesis system required 3–4 months for the regeneration of B.platyphalla plantlets from immature zygotic embryos.
基金This work was supported by Jilin Provincial Natural Science Foundation Project[20190201168JC]“Thirteenth Five-Year”Science and Technology Project of Jilin Provincial Department of Education[JJKH20200341KJ]+3 种基金Science and Technology Project of Jilin Provincial Department of Education[JJKH20210351KJ,JJKH20210346KJ]Jilin Province Science and Technology Development Plan Project[20200402023NC]Jilin Province Key Technology R&D Project[20180201029NY]Jilin Province Support for the Science and Technology Development Program[20190802012ZG]College Student Innovation and Entrepreneurship Training Program Project[202010193087].Furthermore,the funding body have no role in designing of study,collection and interpretation of data or writing up the manuscript.
文摘Immature embryos of inbred maize(Zea mays)lines(H8183,H8184,and H8185)were used for Agrobacterium infection.We used theβ-glucuronidase gene(GUS)as the target gene and the glufosinate resistance gene(bar)as the selection marker.We conducted research on several aspects,such as different genotypes,coculture conditions,screening agent concentrations,and concentrations of indole-3-butytric acid(IBA),6-benzylaminopurine(6-BA),and ascorbic acid(Vc)in the differentiation medium.We optimized the genetic transformation system,and the obtained results indicated that among the three lines studied,the induction rate of H8185 was the highest at 93.2%,followed by H8184,with H8183 having the lowest induction rate(80.1%).The best coculture method was that using the N6 coculture medium layered with a sterile filter paper.Using orthogonal analysis,we found that the optimal combination of the three factors in the differentiation medium was A_(3)(1 mg mL^(−1)IBA),B_(3)C_(1)(1.6 mg mL^(−1)6-BA),and D_(3)(1.5 mg mL^(−1)Vc).Through GUS staining analysis,Bar test-strip analysis,and polymerase chain reaction,five transgenic plants were finally obtained.This study established the optimal conditions for genetic transformation in maize.