The coding sequence of Vitreoscilla hemoglobin (vhb) was cloned with PCR technique from Vitreoscilla stercoraria Pringsheim. The plant expression vector with vhb gene under the control of CaMV 35S promoter was constru...The coding sequence of Vitreoscilla hemoglobin (vhb) was cloned with PCR technique from Vitreoscilla stercoraria Pringsheim. The plant expression vector with vhb gene under the control of CaMV 35S promoter was constructed and used in the transformation of Petunia hybrida Vilm by the Agrobacterium mediated procedure. The results of PCR amplification and Southern hybridization indicated that the vhb gene had been integrated into the petunia genome and the vhb gene expression had been detected by RT-PCR amplification. In hydroponic culture the transgenic petunias grew much better than non-transgenic controls. For further analysis of hypoxia tolerance of transgenic petunia, the petunia plants with vhb gene were submerged into liquid MS medium. The transgenic plants survived in hypoxic condition and grew out of the liquid surface in a few weeks, while non-transgenic plants were still submerged and suffocated in culture solution without ability to grow out of liquid medium in submersed culture for four to five weeks. The vhb gene transformed petunia plants had been planted and tested in a simulated flooding condition, and showed obvious tolerance to water-logging. It seen is that hemoglobin gene from Vitreoscilla might have the potential use in molecular breeding for the improvement of plant resistance to hypoxia and flooding.展开更多
The inheritance of plastid DNA in Pharbitis was studied by the method of restriction fragment length polymorphisms (RFLP).Experimental results showed that plastid DNA from Pharbitis was paternally inherited in recipro...The inheritance of plastid DNA in Pharbitis was studied by the method of restriction fragment length polymorphisms (RFLP).Experimental results showed that plastid DNA from Pharbitis was paternally inherited in reciprocal crosses,P. nil ×P. limbata and P. limbata×P. nil hybrids.But,in the cross of P. limbata×P. nil,the possibility of biparental inheritance of plastid DNA could not be roled out in our preliminary experiment.Thus Pharbitis became the third genus among angiosperms characterized with male plastid transmission.The mechanisms of paternal plastids DNA inheritance in Pharbitis is unclear.The authors proposed that dilution,exclusion and/or degeneration of maternal plastid,including their DNA,after fertilization should be considered.展开更多
[Objective]The research aimed at investigating mechanisms of the changes of pharbitis.nil flower colors and leaf colors.[Method]The recombinant plasmid pMD18-T/Tpn was constructed by amplifying the target fragments fr...[Objective]The research aimed at investigating mechanisms of the changes of pharbitis.nil flower colors and leaf colors.[Method]The recombinant plasmid pMD18-T/Tpn was constructed by amplifying the target fragments from young leaf tissues of the blue and white mutant of Pharbitis nil with PCR technology and investigating in the fields.[Result]The results of investigations in the fields suggested that the mutant was segregated in F3 generation,and the mutations of leaf colors did not affect the changes of flower colors.The results of PCR amplification showed that there was a band about 330 bp in genomic DNA of Lanbai,Chunlan,Dahua,respectively,however,no bands were found in genomic DNA of Chunguang,Baixue,Ping＇anhong and Chuiman.[Conclusion]The sequence of recombinant plasmid of the mutant was indeed a fragment of the transposon of Pharbitis nil.展开更多
In this study,the seeds of wild Petunia Juss.were used as explants to investigate the optimal condition for tissue culture.Several different kinds and concentrations of growth regulators were adopted to produce more m...In this study,the seeds of wild Petunia Juss.were used as explants to investigate the optimal condition for tissue culture.Several different kinds and concentrations of growth regulators were adopted to produce more multiple bud clumps,callus or roots in this study.The experiments may provide experimental foundation for the rapid propagation technology and establishment of tissue culture system for wild Petunia Juss.展开更多
It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth...It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth. To elucidate whether plant hormones are involved in these processes, the effects of exogenous phytohormones, indole-3-acetic acid （IAA）, abscisic acid （ABA）, gibberellin A3 （GA3） and cytokinin （kinetin） on the growth, PM polarization, actin cytoskeleton （AC） organization and cytoplasmic pH （pile） of in vitro 4 h-growing petunia pollen tubes were investigated. IAA, ABA and GA3 displayed the growth-stimulating effects and these were accompanied by orthovanadate-sensitive hyperpolarization of the PM. Fluorescent labeling the enzyme with H＋-ATPase antibodies exhibited IAA- and ABA-induced lateral PM redistribution of it into the subapical zone of pollen tube PM. Pollen cultivation on the medium with latrunculin B, the inhibitor of actin polymerization, resulted in inhibition of pollen tube growth and simultaneously in the drop of endogenous IAA content. The IAA-growth stimulating effect was correlated with increased content of actin filaments （AF） in both apical and subapical zones of tubes, while ABA and GA3 exerted the same effect but it was accompanied by redistributing F-actin only to apical zone. In contrast, kinetin decreased the total F-actin content and inhibited pollen tube growth. It has been shown that the pHe of growing pollen tubes is sensitive to the plant hormones. In the case of male gametophyte growing for 1, 2 and 4 h, IAA induced alkalinization of the cytosol, while ABA and GA3 exerted qualitatively similar effect only after its growth for 1 h and 4 h, respectively. Kinetin, in contrast, resulted in acidification of the cytosol. All these results, taken together, indicate, for the first time, potential targets of the phytohormone action in pollen tubes.展开更多
文摘The coding sequence of Vitreoscilla hemoglobin (vhb) was cloned with PCR technique from Vitreoscilla stercoraria Pringsheim. The plant expression vector with vhb gene under the control of CaMV 35S promoter was constructed and used in the transformation of Petunia hybrida Vilm by the Agrobacterium mediated procedure. The results of PCR amplification and Southern hybridization indicated that the vhb gene had been integrated into the petunia genome and the vhb gene expression had been detected by RT-PCR amplification. In hydroponic culture the transgenic petunias grew much better than non-transgenic controls. For further analysis of hypoxia tolerance of transgenic petunia, the petunia plants with vhb gene were submerged into liquid MS medium. The transgenic plants survived in hypoxic condition and grew out of the liquid surface in a few weeks, while non-transgenic plants were still submerged and suffocated in culture solution without ability to grow out of liquid medium in submersed culture for four to five weeks. The vhb gene transformed petunia plants had been planted and tested in a simulated flooding condition, and showed obvious tolerance to water-logging. It seen is that hemoglobin gene from Vitreoscilla might have the potential use in molecular breeding for the improvement of plant resistance to hypoxia and flooding.
文摘The inheritance of plastid DNA in Pharbitis was studied by the method of restriction fragment length polymorphisms (RFLP).Experimental results showed that plastid DNA from Pharbitis was paternally inherited in reciprocal crosses,P. nil ×P. limbata and P. limbata×P. nil hybrids.But,in the cross of P. limbata×P. nil,the possibility of biparental inheritance of plastid DNA could not be roled out in our preliminary experiment.Thus Pharbitis became the third genus among angiosperms characterized with male plastid transmission.The mechanisms of paternal plastids DNA inheritance in Pharbitis is unclear.The authors proposed that dilution,exclusion and/or degeneration of maternal plastid,including their DNA,after fertilization should be considered.
文摘[Objective]The research aimed at investigating mechanisms of the changes of pharbitis.nil flower colors and leaf colors.[Method]The recombinant plasmid pMD18-T/Tpn was constructed by amplifying the target fragments from young leaf tissues of the blue and white mutant of Pharbitis nil with PCR technology and investigating in the fields.[Result]The results of investigations in the fields suggested that the mutant was segregated in F3 generation,and the mutations of leaf colors did not affect the changes of flower colors.The results of PCR amplification showed that there was a band about 330 bp in genomic DNA of Lanbai,Chunlan,Dahua,respectively,however,no bands were found in genomic DNA of Chunguang,Baixue,Ping＇anhong and Chuiman.[Conclusion]The sequence of recombinant plasmid of the mutant was indeed a fragment of the transposon of Pharbitis nil.
基金Supported by Talent Introduction Project of Anhui Science and Technology University(ZRC2007147)General Project of Natural Scientific Research from Education Department of Anhui Province(KJ2010B052,KJ2010B294)
文摘In this study,the seeds of wild Petunia Juss.were used as explants to investigate the optimal condition for tissue culture.Several different kinds and concentrations of growth regulators were adopted to produce more multiple bud clumps,callus or roots in this study.The experiments may provide experimental foundation for the rapid propagation technology and establishment of tissue culture system for wild Petunia Juss.
文摘It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth. To elucidate whether plant hormones are involved in these processes, the effects of exogenous phytohormones, indole-3-acetic acid （IAA）, abscisic acid （ABA）, gibberellin A3 （GA3） and cytokinin （kinetin） on the growth, PM polarization, actin cytoskeleton （AC） organization and cytoplasmic pH （pile） of in vitro 4 h-growing petunia pollen tubes were investigated. IAA, ABA and GA3 displayed the growth-stimulating effects and these were accompanied by orthovanadate-sensitive hyperpolarization of the PM. Fluorescent labeling the enzyme with H＋-ATPase antibodies exhibited IAA- and ABA-induced lateral PM redistribution of it into the subapical zone of pollen tube PM. Pollen cultivation on the medium with latrunculin B, the inhibitor of actin polymerization, resulted in inhibition of pollen tube growth and simultaneously in the drop of endogenous IAA content. The IAA-growth stimulating effect was correlated with increased content of actin filaments （AF） in both apical and subapical zones of tubes, while ABA and GA3 exerted the same effect but it was accompanied by redistributing F-actin only to apical zone. In contrast, kinetin decreased the total F-actin content and inhibited pollen tube growth. It has been shown that the pHe of growing pollen tubes is sensitive to the plant hormones. In the case of male gametophyte growing for 1, 2 and 4 h, IAA induced alkalinization of the cytosol, while ABA and GA3 exerted qualitatively similar effect only after its growth for 1 h and 4 h, respectively. Kinetin, in contrast, resulted in acidification of the cytosol. All these results, taken together, indicate, for the first time, potential targets of the phytohormone action in pollen tubes.
