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
Previous studies have shown that high light intensity can induce anthocyanin synthesis(AS)in petunia plants.To identifywhich kind of light quality plays a role in inducing such metabolic process,and what transcripts p...Previous studies have shown that high light intensity can induce anthocyanin synthesis(AS)in petunia plants.To identifywhich kind of light quality plays a role in inducing such metabolic process,and what transcripts participate in controlling it,we carried out whole-transcriptome sequencing and analysis of petunia petals treated with different light-quality conditions.Among the red and white light treatments,a total of 2205 differentially expressed genes and 15,22,and 20 differentially expressed circRNAs,miRNAs,and lncRNAs,were identified respectively.The AS-related genes,including the structural genes CHSj,F3H,F35H,DFR,and ANS,and the regulatory genes AN4,DPL,PHZ and MYBx were found to be downregulated under red light condition compared with their levels under white light condition.Furthermore,the light photoreceptor Cryptochrome 3(CRY3)and a series of light-dependent genes,such as PIF,HY5,andBBXs,were also determined to respond to the light treatments.The anthocyanin contents in early petunia petals under red light were significantly lower than that under white and blue light.The results of qRT-PCR further confirmed the expression pattern of some AS-related and light-response genes in response to different light quality.Yeast two-hybrid results showed that the key elements in the light signal pathway,HY5 can interact with BBX19,BBX24 and BBX25.And PHZ,the important AS regulator can induce anthocyanin synthesis in response to blue light quality fromtransient expression analysis in petunia petals.These findings presented here not only deepen our understanding of how light quality controls anthocyanin synthesis,but also allow us to explore potential target genes for improving pigment production in petunia flower petals.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
Salinity is one of the abiotic stresses that limits the growth and productivity of many crops.A possible survival strategy for plant under saline conditions is to use compounds that could minimize the harmful effects ...Salinity is one of the abiotic stresses that limits the growth and productivity of many crops.A possible survival strategy for plant under saline conditions is to use compounds that could minimize the harmful effects of salt stress on the plant development.The objective of the presented study was to investigate the effect of exogenous ascorbic acid(ASA)with or without gibberellic acid(GA3)on key growth and biochemical parameters in two petunia cultivars‘Prism Rose’and‘Prism White’under saline(150 mM NaCl)and non-saline in vitro condition.Nodal cutting with an axillary buds were used as explants.Application of 1 mM ascorbic acid with or without 0.05 mM gibberellic acid into the MS medium stimulated the length of shoots and the number of new shoots of‘Prism Rose’;whereas,it decreased the root length and the number of roots of both‘Prism Rose’and‘Prism White’under non-saline condition.The addition of ascorbic acid with or without gibberellic acid into the MS medium under saline condition,increased the length of plants and the number of new shoots,but did not affect their root number and length.NaCl treatments increased the proline content and lipid peroxidation which was indicated by the accumulation of malondialdehyde(MDA).The study revealed a correlation between chlorophylls a and b content and the leaf pigmentation intensity–parameter a*.Addition of 1 mM ascorbic acid with 0.05 mM gibberellic acid into the MS medium plays a protective role in salinity tolerance by improving the shoot growth and the development as well as increasing the activities of the antioxidant enzymes and other antioxidant substances.展开更多
As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a ...As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a necessary element for plant growth, and soluble silicon can improve plant resilience. To improve the drought resilience of Petunia hybrida, the silicon transporter protein OsLsi1 and OsLsi2 genes cloned from rice（Oryza sative） were transferred into Petunia hybrida by Agrobacterium-mediated method, and finally got 26 and 32 positive plants, respectively by PCR and RT-PCR detections. With a control of non-transgenic plants, the obtained transgenic plants were taken by drought treatment stress for 0, 4, 7, 10 and 14 days, then re-watered and measured physiological indexes as malondialdehyde（MDA） content, free proline（Pro） content, superoxide dismutase（SOD） activity and peroxidase（POD） activity to study the effect of Petunia＇s drought resistance. All the results proved that the silicon transporter protein OsLsi1 and OsLsi2 genes were normally transcripted and expressed in transgenic Petunia hybrida; OsLsi1 gene could improve the abilities of plants＇ drought resistance and recover after drought stress, while OsLsi2 gene could reduce the above abilities. The order of the drought resistance ability of the three strains from strong to weak was OsLsi1〉CK〉OsLsi2; and silicon indeed improved the ability of drought resistance as well. All these results provided a new way to improve the drought resistance of Petunia, and laid a foundation to improve the ability of garden plants＇ drought resistance and water saving.展开更多
Water insufficiency is the hampering feature of crop sustainability,especially in arid and semi-arid regions.So,the effectual usage of all water resources especially underground brackish water represents the core prio...Water insufficiency is the hampering feature of crop sustainability,especially in arid and semi-arid regions.So,the effectual usage of all water resources especially underground brackish water represents the core priority in Saudi Arabia.The present study aimed to recognize the influence of different types of water irrigation(tap water as a control,salinized well water,and magnetized salinized well water)with or without soil amendments(soil without any amendment as a control,peat-moss,ferrous sulfate,and peat-moss plus ferrous sulfate)on petunia plant growth and flowering as well as ion content.Irrigating Petunia plants with saline well water adversely affected growth and flowering as compared to tap water and magnetized saline well water.Additionally,plants irrigated with magnetized water showed a significant enhancement in all the studied vegetative and flowering growth parameters as compared to those irrigated with salinized well water.Furthermore,mineral contents and survival of Petunia plants irrigated with magnetized well water were higher than those irrigated with tap water.Irrigation with magnetized well water significantly reduced levels of Na+and Cl−ions in leaves of Petunia plants indicating the role of magnetization in alleviating harmful effects of salinity.In conclusion,we recommend the utilization of magnetized saline well water for irrigating Petunia plants either alone or in combination with soil amendments(peat moss plus ferrous sulfate).展开更多
The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local c...The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local conditions.The experiment included four treatments with three kinds of forliar fertilizers—Dau Trau MK 30-10-5,gibberellin 25-10-10 and abscisic acid.The control treatment(T0)used sterilized water without foliar fertilizer.The results showed that all the foliar fertilizers influenced well on growth and development of Petunia hybrida.Dau Trau MK 30-10-5 helped Petunia hybrida have high quality and high value/cost ratio than the others.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
Self-Incompatibility （SI） Is a genetic mechanism of self/non-self pollen recognition to prevent self-fertilization In many flowering plants and, In most cases, this is controlled by a multl-allellc S-locus. S-RNase ...Self-Incompatibility （SI） Is a genetic mechanism of self/non-self pollen recognition to prevent self-fertilization In many flowering plants and, In most cases, this is controlled by a multl-allellc S-locus. S-RNase and Slocus F box （SLF） proteins have been shown to be the female and male determinants of gametophytlc selfIncompatibility （GSI）, respectively, In the Solanaceae, Scrophulariaceae and Rosaceae. Nevertheless, It is thought that additional factors are required for the SI response. Herein, we constructed a mature anther cDNA library from a self-Incompatible Petunia hybrida Vllm. line of the S3S3 haplotype. Using AhS2-RNase from Antirrhinum hispanicum as a bait for yeast two-hybrid screening, we found that petunia germinating pollen （PGP） S/D3 was capable of Interacting physically with the bait. However, the Interaction lacked haplotype specificity. The PGPS/D3 gene Is a single copy gene that Is expressed In tissues such as the style, ovary, pollen, and leaf. The PGPS/D3：：GFP （green fluorescence protein） construct was detected In both the membrane and cytoplasm. The Implications of these findings In the operation of S-RNase-based SI are discussed.展开更多
A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method medi...A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method mediated by Agrobacterium. A series of novel phenotypes was observed in transgenic petunia plants, including the formation of ectopic spikes on the adaxial surface of corollas and small petals on the abaxial surface of corollas, fusion of floral organs, shortening of corolla midribs, the formation of tumor-like knots along the midrib on the abaxial surface and serrated lobs of corolla margins, and alterations in petal color; except for changes in the leaves and plant architecture, RT-PCR showed that the Pttknl gene was expressed in the leaves of different petunia transgenic plants, whereas no signal was detected in wild-type plants. The possible function of Pttknl in leaf and flower development is discussed.展开更多
Chalcone synthase A is a key enzyme in the anthocyanin biosynthesis pathway. Expression of chsA gene in transgenic Petunia hybrida resulted in flower color alterations and co-suppression of transgenes and endogenous g...Chalcone synthase A is a key enzyme in the anthocyanin biosynthesis pathway. Expression of chsA gene in transgenic Petunia hybrida resulted in flower color alterations and co-suppression of transgenes and endogenous genes. We fused the β-glucuronidase (uidA) gene to the C-terminal of chsA gene, and transferred the fusion gene into Petunia hybrida via Agrobacterium tumefaciens. GUS histochemical staining analysis showed that co-suppression occurred specifically during the development of flowers and co-suppression required the mutual interaction of endogenous genes and transgenes. RNA in situ hybridization analysis suggested that co-suppression occurred in the entire plant, and RNA degradation occurred in the cytoplasm.展开更多
Cytochrome P450 CYP2E1 is mainly present in hepatocytes in the livers of mammals,where it plays an important role in the metabolism of xenobiotic organic substances. Previous studies showed that transgenic petunia(Pet...Cytochrome P450 CYP2E1 is mainly present in hepatocytes in the livers of mammals,where it plays an important role in the metabolism of xenobiotic organic substances. Previous studies showed that transgenic petunia(Petunia hybrid) plants harboring a mammalian cyp2e1 gene(designated cyp2e1-transgenic petunia) exhibited increased resistance to formaldehyde stress. In this study,we used cyp2e1-transgenic petunia plants to analyze physiological indexes related to formaldehyde stress responses. The results indicated that under formaldehyde stress,the malondialdehyde content in cyp2e1-transgenic petunia plants was lower than in β-glucuronidase gene(gus)-transgenic and wild-type petunia plants. The activities of both superoxide dismutase and peroxidase in the cyp2e1-transgenic plants were higher than in gus-transgenic and wild-type plants. The alcohol dehydrogenase activity was slightly increased and more glutathione was consumed. Additionally,under formaldehyde stress,the levels of plant hormones including indole-3-acetic acid,zeatin and abscisic acid in cyp2e1-transgenic petunia plants displayed decreasing trends,whereas the level of gibberellic acid displayed an increasing trend. In contrast,the indole-3-acetic acid,zeatin and abscisic acid levels in gus-transgenic and wild-type petunia plants displayed increasing trends,whereas the gibberellic acid level displayed a decreasing trend. At 72 h after incubation of 0.5 g of cyp2e1-transgenic petunia plants in 40 mL of treatment solution containing formaldehyde at 50 mg·L^(-1),the formaldehyde content remaining in the treatment solution was close to zero while approximately half of original formaldehyde remained in the treatment solutions containing gus-transgenic and wild-type petunia plants.展开更多
Virus-induced gene silencing offers a powerful reverse-genetic tool for the study of gene function in plants. We have previously reported ef- fective gene silencing of plant genes using a viral satellite DNA associate...Virus-induced gene silencing offers a powerful reverse-genetic tool for the study of gene function in plants. We have previously reported ef- fective gene silencing of plant genes using a viral satellite DNA associated with Tomato yellow leaf curl China virus (TYLCCNV). In this study,we further modified the viral satellite DNA-based vector. The modified vector can induce sulfu (Su) gene silencing as effective as the original vector in Nicotiana ben- thamiana plants,but the new system simplifies pro- cedures for construction of vector derivative. Fur- thermore,a fragment of petunia Su or chalcone syn- thase (CHS) endogenous gene was inserted into the modified vector. When petunia plants were agro- inoculated with the modified vector carrying a Su or CHS gene,the Su silenced plants started to appear yellowing in veins of systemically infected upper leaves two weeks after agroinoculation,while the CHS silenced plants started to show flower color change one month after agroinoculation and later single-color flowers became mosaic.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.
基金This research was supported by the National Natural Science Foundation of China(Grant No.U1504320)Financial Project of Henan Province(Grant No.2019ZC23)We thank Liwen Bianji,Edanz Group China(www.liwenbianji.cn/ac),for editing the English text of a draft of this manuscript.
文摘Previous studies have shown that high light intensity can induce anthocyanin synthesis(AS)in petunia plants.To identifywhich kind of light quality plays a role in inducing such metabolic process,and what transcripts participate in controlling it,we carried out whole-transcriptome sequencing and analysis of petunia petals treated with different light-quality conditions.Among the red and white light treatments,a total of 2205 differentially expressed genes and 15,22,and 20 differentially expressed circRNAs,miRNAs,and lncRNAs,were identified respectively.The AS-related genes,including the structural genes CHSj,F3H,F35H,DFR,and ANS,and the regulatory genes AN4,DPL,PHZ and MYBx were found to be downregulated under red light condition compared with their levels under white light condition.Furthermore,the light photoreceptor Cryptochrome 3(CRY3)and a series of light-dependent genes,such as PIF,HY5,andBBXs,were also determined to respond to the light treatments.The anthocyanin contents in early petunia petals under red light were significantly lower than that under white and blue light.The results of qRT-PCR further confirmed the expression pattern of some AS-related and light-response genes in response to different light quality.Yeast two-hybrid results showed that the key elements in the light signal pathway,HY5 can interact with BBX19,BBX24 and BBX25.And PHZ,the important AS regulator can induce anthocyanin synthesis in response to blue light quality fromtransient expression analysis in petunia petals.These findings presented here not only deepen our understanding of how light quality controls anthocyanin synthesis,but also allow us to explore potential target genes for improving pigment production in petunia flower petals.
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Salinity is one of the abiotic stresses that limits the growth and productivity of many crops.A possible survival strategy for plant under saline conditions is to use compounds that could minimize the harmful effects of salt stress on the plant development.The objective of the presented study was to investigate the effect of exogenous ascorbic acid(ASA)with or without gibberellic acid(GA3)on key growth and biochemical parameters in two petunia cultivars‘Prism Rose’and‘Prism White’under saline(150 mM NaCl)and non-saline in vitro condition.Nodal cutting with an axillary buds were used as explants.Application of 1 mM ascorbic acid with or without 0.05 mM gibberellic acid into the MS medium stimulated the length of shoots and the number of new shoots of‘Prism Rose’;whereas,it decreased the root length and the number of roots of both‘Prism Rose’and‘Prism White’under non-saline condition.The addition of ascorbic acid with or without gibberellic acid into the MS medium under saline condition,increased the length of plants and the number of new shoots,but did not affect their root number and length.NaCl treatments increased the proline content and lipid peroxidation which was indicated by the accumulation of malondialdehyde(MDA).The study revealed a correlation between chlorophylls a and b content and the leaf pigmentation intensity–parameter a*.Addition of 1 mM ascorbic acid with 0.05 mM gibberellic acid into the MS medium plays a protective role in salinity tolerance by improving the shoot growth and the development as well as increasing the activities of the antioxidant enzymes and other antioxidant substances.
基金Supported by the Fund of Science and Technology Research Project of Education Department in Heilongjiang Province(12531014)
文摘As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a necessary element for plant growth, and soluble silicon can improve plant resilience. To improve the drought resilience of Petunia hybrida, the silicon transporter protein OsLsi1 and OsLsi2 genes cloned from rice（Oryza sative） were transferred into Petunia hybrida by Agrobacterium-mediated method, and finally got 26 and 32 positive plants, respectively by PCR and RT-PCR detections. With a control of non-transgenic plants, the obtained transgenic plants were taken by drought treatment stress for 0, 4, 7, 10 and 14 days, then re-watered and measured physiological indexes as malondialdehyde（MDA） content, free proline（Pro） content, superoxide dismutase（SOD） activity and peroxidase（POD） activity to study the effect of Petunia＇s drought resistance. All the results proved that the silicon transporter protein OsLsi1 and OsLsi2 genes were normally transcripted and expressed in transgenic Petunia hybrida; OsLsi1 gene could improve the abilities of plants＇ drought resistance and recover after drought stress, while OsLsi2 gene could reduce the above abilities. The order of the drought resistance ability of the three strains from strong to weak was OsLsi1〉CK〉OsLsi2; and silicon indeed improved the ability of drought resistance as well. All these results provided a new way to improve the drought resistance of Petunia, and laid a foundation to improve the ability of garden plants＇ drought resistance and water saving.
基金funded by Abdulaziz City for Science and Technology,Saudi Arabia(Grant Research No.1-17-04-001-0021).
文摘Water insufficiency is the hampering feature of crop sustainability,especially in arid and semi-arid regions.So,the effectual usage of all water resources especially underground brackish water represents the core priority in Saudi Arabia.The present study aimed to recognize the influence of different types of water irrigation(tap water as a control,salinized well water,and magnetized salinized well water)with or without soil amendments(soil without any amendment as a control,peat-moss,ferrous sulfate,and peat-moss plus ferrous sulfate)on petunia plant growth and flowering as well as ion content.Irrigating Petunia plants with saline well water adversely affected growth and flowering as compared to tap water and magnetized saline well water.Additionally,plants irrigated with magnetized water showed a significant enhancement in all the studied vegetative and flowering growth parameters as compared to those irrigated with salinized well water.Furthermore,mineral contents and survival of Petunia plants irrigated with magnetized well water were higher than those irrigated with tap water.Irrigation with magnetized well water significantly reduced levels of Na+and Cl−ions in leaves of Petunia plants indicating the role of magnetization in alleviating harmful effects of salinity.In conclusion,we recommend the utilization of magnetized saline well water for irrigating Petunia plants either alone or in combination with soil amendments(peat moss plus ferrous sulfate).
文摘The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local conditions.The experiment included four treatments with three kinds of forliar fertilizers—Dau Trau MK 30-10-5,gibberellin 25-10-10 and abscisic acid.The control treatment(T0)used sterilized water without foliar fertilizer.The results showed that all the foliar fertilizers influenced well on growth and development of Petunia hybrida.Dau Trau MK 30-10-5 helped Petunia hybrida have high quality and high value/cost ratio than the others.
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.
基金Supported by the Chinese Academy of Sciences and the National Natural Science Foundation of China （30221002）.Acknowledgements The authors are grateful to Qi Xie （Institute of Genetics and Developmental Biology, the Chinese Academy of Sciences） for help with the construction of the cDNA library and the yeast two-hybrid techniques and Tim Robbins for providing P. hybrida. The authors also thank Jiayang Li （Institute of Genetics and Developmental Biology, the Chinese Academy of Sciences） for the pB1221-35S：GFP vector.
文摘Self-Incompatibility （SI） Is a genetic mechanism of self/non-self pollen recognition to prevent self-fertilization In many flowering plants and, In most cases, this is controlled by a multl-allellc S-locus. S-RNase and Slocus F box （SLF） proteins have been shown to be the female and male determinants of gametophytlc selfIncompatibility （GSI）, respectively, In the Solanaceae, Scrophulariaceae and Rosaceae. Nevertheless, It is thought that additional factors are required for the SI response. Herein, we constructed a mature anther cDNA library from a self-Incompatible Petunia hybrida Vllm. line of the S3S3 haplotype. Using AhS2-RNase from Antirrhinum hispanicum as a bait for yeast two-hybrid screening, we found that petunia germinating pollen （PGP） S/D3 was capable of Interacting physically with the bait. However, the Interaction lacked haplotype specificity. The PGPS/D3 gene Is a single copy gene that Is expressed In tissues such as the style, ovary, pollen, and leaf. The PGPS/D3：：GFP （green fluorescence protein） construct was detected In both the membrane and cytoplasm. The Implications of these findings In the operation of S-RNase-based SI are discussed.
基金Supported by the Gansu Provincial Natural Science Foundation of China （ZS021-A25-047-N） and Xi＇an Urban Rural Construction Committee.Acknowledgements The authors thank Dr. 0lof 0lsson （Goeteberg University, Goeteberg, Sweden） for providing the plasmid containing 35S：：Pttkn1.
文摘A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method mediated by Agrobacterium. A series of novel phenotypes was observed in transgenic petunia plants, including the formation of ectopic spikes on the adaxial surface of corollas and small petals on the abaxial surface of corollas, fusion of floral organs, shortening of corolla midribs, the formation of tumor-like knots along the midrib on the abaxial surface and serrated lobs of corolla margins, and alterations in petal color; except for changes in the leaves and plant architecture, RT-PCR showed that the Pttknl gene was expressed in the leaves of different petunia transgenic plants, whereas no signal was detected in wild-type plants. The possible function of Pttknl in leaf and flower development is discussed.
基金the National Natural Science Foundation of China (Grant No. 39670415), Yunnan Province-Peking University joint project (B9810K) and Shenzhen Science and Technology Bureau.
文摘Chalcone synthase A is a key enzyme in the anthocyanin biosynthesis pathway. Expression of chsA gene in transgenic Petunia hybrida resulted in flower color alterations and co-suppression of transgenes and endogenous genes. We fused the β-glucuronidase (uidA) gene to the C-terminal of chsA gene, and transferred the fusion gene into Petunia hybrida via Agrobacterium tumefaciens. GUS histochemical staining analysis showed that co-suppression occurred specifically during the development of flowers and co-suppression required the mutual interaction of endogenous genes and transgenes. RNA in situ hybridization analysis suggested that co-suppression occurred in the entire plant, and RNA degradation occurred in the cytoplasm.
基金supported by the Hangzhou Science and Technology Development Plan(Grant No.20140432B05)the New-shoot Talents Program of Zhejiang Province(Grant No.2013R421008)
文摘Cytochrome P450 CYP2E1 is mainly present in hepatocytes in the livers of mammals,where it plays an important role in the metabolism of xenobiotic organic substances. Previous studies showed that transgenic petunia(Petunia hybrid) plants harboring a mammalian cyp2e1 gene(designated cyp2e1-transgenic petunia) exhibited increased resistance to formaldehyde stress. In this study,we used cyp2e1-transgenic petunia plants to analyze physiological indexes related to formaldehyde stress responses. The results indicated that under formaldehyde stress,the malondialdehyde content in cyp2e1-transgenic petunia plants was lower than in β-glucuronidase gene(gus)-transgenic and wild-type petunia plants. The activities of both superoxide dismutase and peroxidase in the cyp2e1-transgenic plants were higher than in gus-transgenic and wild-type plants. The alcohol dehydrogenase activity was slightly increased and more glutathione was consumed. Additionally,under formaldehyde stress,the levels of plant hormones including indole-3-acetic acid,zeatin and abscisic acid in cyp2e1-transgenic petunia plants displayed decreasing trends,whereas the level of gibberellic acid displayed an increasing trend. In contrast,the indole-3-acetic acid,zeatin and abscisic acid levels in gus-transgenic and wild-type petunia plants displayed increasing trends,whereas the gibberellic acid level displayed a decreasing trend. At 72 h after incubation of 0.5 g of cyp2e1-transgenic petunia plants in 40 mL of treatment solution containing formaldehyde at 50 mg·L^(-1),the formaldehyde content remaining in the treatment solution was close to zero while approximately half of original formaldehyde remained in the treatment solutions containing gus-transgenic and wild-type petunia plants.
基金the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China （Grant No. 705025） the National Key Basic Research and Developmenl Program （Grant No. 2006CBI01903） the National Natural Science Foundation of China （Grants Nos, 30301H94 and 30530520）.
文摘Virus-induced gene silencing offers a powerful reverse-genetic tool for the study of gene function in plants. We have previously reported ef- fective gene silencing of plant genes using a viral satellite DNA associated with Tomato yellow leaf curl China virus (TYLCCNV). In this study,we further modified the viral satellite DNA-based vector. The modified vector can induce sulfu (Su) gene silencing as effective as the original vector in Nicotiana ben- thamiana plants,but the new system simplifies pro- cedures for construction of vector derivative. Fur- thermore,a fragment of petunia Su or chalcone syn- thase (CHS) endogenous gene was inserted into the modified vector. When petunia plants were agro- inoculated with the modified vector carrying a Su or CHS gene,the Su silenced plants started to appear yellowing in veins of systemically infected upper leaves two weeks after agroinoculation,while the CHS silenced plants started to show flower color change one month after agroinoculation and later single-color flowers became mosaic.
