An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorop...An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.展开更多
Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles ...Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles and techniques of plant propagation and improvement. Microstructures and fluctuations of phytohormones in the adventitious rooting were studied with the etiolated soft- wood shoots of Paeonia suffkuticosa 'Yinfen Jinlin'. There are no pre-primordia in the shoots of the cultivar. Adventitious roots are produced in five stages: shoot selection, primordium initiation, primordium growth, conducting tissue differentiation and root protru- sion. Primordia initiated in the cortex. The contents of the endogenous hormones, IAA, ABA and GA, were 5.842, 0.873 and 1.043 nmol·g^-1 FW on the bases of shoots, respectively. CTKs which included isopentenyl adenine (iPA), zeatin riboside (ZR) and dihy- drozeatin riboside (DHZR) were 0.949, 0.695 and 2.034 nmol·g^-1 FW, respectively. DHZR is active among CTKs. The ratio of IAA to GA, CTK and ABA clearly increased at the stage of primordium initiation, while they showed low levels at the stages of primor- dium growth. The ratios were restored at the shoot levels at the stage of root protrusion. IBA provoked primordia initiation in the cortex, the vascular cambium, the pith and even in the callus induced on the base of shoots. ]AA levels in the treated shoots increased gradually to its highest level (three times of control) at the stage of conducting tissue differentiation. The ratios of IAA to GA, CTK and ABA clearly decreased at the stage of primordium initiation. The ratio of IAA to ABA is regulated at 10:1.展开更多
The effect of stock-plant etiolation on adventitious rooting of single-node leafy cuttings (SNCs) made from coppice shoots from different age groups of donor plants were investigated for teak (Tectona grandis Linn. f....The effect of stock-plant etiolation on adventitious rooting of single-node leafy cuttings (SNCs) made from coppice shoots from different age groups of donor plants were investigated for teak (Tectona grandis Linn. f.). When donor plants age were 1-, 2-, 3-, 4-, and 5-year-old, they were coppiced and maintained in the dark for etiolation, while a parallel set was kept in normal light in an open environment. After 20 days, coppice shoots were made into SNCs which were cultured under intermittent mist for rooting. Stock-plant etiolation significantly increased percent rooting, shoot and root length, and number of roots per SNC, but callusing at the base of SNCs decreased. Etiolated SNCs have shown 71.7% rooting, whereas for controls (non-etiolated), the response was 41.7%. The effect of aging on callusing, rooting and sprouting of etiolated and non-etiolated coppice shoots cuttings varied widely. Aging of donor plants decreased rooting and sprouting capability in SNCs and increased callusing at the base of etiolated cuttings. The results showed that stock-plant etiolation in 1-, 2-, 3-, 4-, and 5-year-old donor plants hastened rooting potential by rejuvenation of coppice shoots.展开更多
The possible nutrient problems existing in leaf etiolation,the influence of film mulching on the yield of peanut and benefits of balanced fertilization were analyzed. The results showed that the deficiency of nutrient...The possible nutrient problems existing in leaf etiolation,the influence of film mulching on the yield of peanut and benefits of balanced fertilization were analyzed. The results showed that the deficiency of nutrient elements was not main limiting factor in leaf etiolation of peanut in Hong'an. Deep trench,film mulching,liming and organic manure could be used to prevent leaf etiolation. The film mulching could increase the yield of peanut by improving the rate of germination and seedling,the number of branches and full pod,and it was beneficial to playing the effect of balanced fertilization.展开更多
Phytochrome-interacting factors (PIFs) regulate an array of developmental responses ranging from seed germi- nation to vegetational architecture in Arabidopsis. However, information regarding the functions of the PI...Phytochrome-interacting factors (PIFs) regulate an array of developmental responses ranging from seed germi- nation to vegetational architecture in Arabidopsis. However, information regarding the functions of the PIF family in monocots has not been widely reported. Here, we investigate the roles of OsPIL15, a member of the rice (Oryza sativa L. cv. Nipponbare) PIF family, in regulating seedling growth. OsPIL15 encodes a basic helix-loop-helix factor localized in the nucleus. OsPIL15-OX seedlings exhibit an exaggerated shorter above- ground part and undeveloped root system relative to wild-type seedlings, suggesting that OsPIL15 represses seedling growth in the dark. Microarray analysis combined with gene ontology analysis revealed that OsPIL15 represses a set of genes involved in auxin pathways and cell wall organization or biogenesis. Given the important roles of the auxin pathway and cell wall properties in controlling plant growth, we speculate that OsPIL15 represses seedling growth likely by regulating the auxin pathway and suppressing cell wall organization in etiolated rice seedlings. Additionally, exposure to red light or far-red light relieved growth retardation and promoted seedling elongation in the OsPIL15-OX lines, despite higher levels of OsPIL15 transcripts under red light and far-red light than in the dark. These results suggest that light regulation of OsPIL15 expres- sion is probably involved in photomorphogenesis in rice.展开更多
Proteomic analysis was performed on seedlings after different light treatments. A total of (1 350±31) protein spots was separated and visualized on each silver nitrate-stained two-dimensional gel using protein ...Proteomic analysis was performed on seedlings after different light treatments. A total of (1 350±31) protein spots was separated and visualized on each silver nitrate-stained two-dimensional gel using protein samples prepared from light-grown or etiolated seedlings with or without 6-9 h light treatment. Twenty-five protein spots (encoded by 19 genes) that were significantly accumulated upon light treatment were identified using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry method. Functional proteomics indicated that these proteins involved mainly in chloroplast development, energy metabolism, cell cycle progression and membrane electron transport. For 18 of the protein-coding genes we identified through an internet search, the transcript levels of 17 genes matched roughly with their protein content in etiolated and green seedlings, suggesting that these genes were regulated by light mainly at the transcriptional level. Despite a very significant increase in the amount of proteins upon light treatment, similar RNA levels were found in dark-grown or green seedlings for the carbonic anhydrase gene At3g05100, indicating a possible post-transcriptional regulatory mechanism. Elucidation of light-induced protein accumulation will undoubtedly enhance our understanding of plant photomorphogenesis.展开更多
Ambient light has profound effects on early seedling de-etiolation through red and far-red light-absorbing phytochromes and blue and UV-A light-absorbing cryptochromes. Subsequent integration of various light signal t...Ambient light has profound effects on early seedling de-etiolation through red and far-red light-absorbing phytochromes and blue and UV-A light-absorbing cryptochromes. Subsequent integration of various light signal trans- duction pathways leads to changes in gene expression and morphogenic responses. Here, we report the isolation of a new Arabidopsis light-signaling component, HYPOSENSITIVE TO LIGHT or HTL. Both htl-1 and htl-2 alleles displayed a long hypocotyl phenotype under red, far-red, and blue light, whereas overexpression of HTL caused a short hypocotyl pheno- type under similar light conditions. The mutants also showed other photomorphogenic defects such as elongated petioles, retarded cotyledon and leaf expansion, reduced accumulation of chlorophyll and anthocyanin pigments, and attenuated expression of light-responsive CHLOROPHYLL A/B BINDING PROTEIN 3 and CHALCONE SYNTHASE genes. HTL belongs to an alpha/beta fold protein family and is localized strongly in the nucleus and weakly in the cytosol. The expression of HTL was strongly induced by light of various wavelengths and this light induction was impaired in elongated hypocotyl 5. HY5 directly bound to both a C/G-box and a G-box in the HTL promoter but with a greater affinity toward the C/G-box. HTL, therefore, represents a new signaling step downstream of HY5 in phy- and cry-mediated de-etiolation responses.展开更多
Soluble invertase was purified from pea (Pisum sativum L.) by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite col...Soluble invertase was purified from pea (Pisum sativum L.) by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid (SAC) and alkaline (SALK) invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.32072049)Open Foundation of State Key Laboratory of Rice Biology,China(Grant No.20210208).
文摘An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.
文摘Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles and techniques of plant propagation and improvement. Microstructures and fluctuations of phytohormones in the adventitious rooting were studied with the etiolated soft- wood shoots of Paeonia suffkuticosa 'Yinfen Jinlin'. There are no pre-primordia in the shoots of the cultivar. Adventitious roots are produced in five stages: shoot selection, primordium initiation, primordium growth, conducting tissue differentiation and root protru- sion. Primordia initiated in the cortex. The contents of the endogenous hormones, IAA, ABA and GA, were 5.842, 0.873 and 1.043 nmol·g^-1 FW on the bases of shoots, respectively. CTKs which included isopentenyl adenine (iPA), zeatin riboside (ZR) and dihy- drozeatin riboside (DHZR) were 0.949, 0.695 and 2.034 nmol·g^-1 FW, respectively. DHZR is active among CTKs. The ratio of IAA to GA, CTK and ABA clearly increased at the stage of primordium initiation, while they showed low levels at the stages of primor- dium growth. The ratios were restored at the shoot levels at the stage of root protrusion. IBA provoked primordia initiation in the cortex, the vascular cambium, the pith and even in the callus induced on the base of shoots. ]AA levels in the treated shoots increased gradually to its highest level (three times of control) at the stage of conducting tissue differentiation. The ratios of IAA to GA, CTK and ABA clearly decreased at the stage of primordium initiation. The ratio of IAA to ABA is regulated at 10:1.
文摘The effect of stock-plant etiolation on adventitious rooting of single-node leafy cuttings (SNCs) made from coppice shoots from different age groups of donor plants were investigated for teak (Tectona grandis Linn. f.). When donor plants age were 1-, 2-, 3-, 4-, and 5-year-old, they were coppiced and maintained in the dark for etiolation, while a parallel set was kept in normal light in an open environment. After 20 days, coppice shoots were made into SNCs which were cultured under intermittent mist for rooting. Stock-plant etiolation significantly increased percent rooting, shoot and root length, and number of roots per SNC, but callusing at the base of SNCs decreased. Etiolated SNCs have shown 71.7% rooting, whereas for controls (non-etiolated), the response was 41.7%. The effect of aging on callusing, rooting and sprouting of etiolated and non-etiolated coppice shoots cuttings varied widely. Aging of donor plants decreased rooting and sprouting capability in SNCs and increased callusing at the base of etiolated cuttings. The results showed that stock-plant etiolation in 1-, 2-, 3-, 4-, and 5-year-old donor plants hastened rooting potential by rejuvenation of coppice shoots.
基金Supported by International Plant Nutrition Institute(IPNI-Hubei-28)Modern Agro-industry Technology Research System(CARS-14)
文摘The possible nutrient problems existing in leaf etiolation,the influence of film mulching on the yield of peanut and benefits of balanced fertilization were analyzed. The results showed that the deficiency of nutrient elements was not main limiting factor in leaf etiolation of peanut in Hong'an. Deep trench,film mulching,liming and organic manure could be used to prevent leaf etiolation. The film mulching could increase the yield of peanut by improving the rate of germination and seedling,the number of branches and full pod,and it was beneficial to playing the effect of balanced fertilization.
基金supported by grants from the National Natural Science Foundation of China (31270232 and 30971744)the Shandong Natural Science Funds for Distinguished Young Scholars (JQ200911)
文摘Phytochrome-interacting factors (PIFs) regulate an array of developmental responses ranging from seed germi- nation to vegetational architecture in Arabidopsis. However, information regarding the functions of the PIF family in monocots has not been widely reported. Here, we investigate the roles of OsPIL15, a member of the rice (Oryza sativa L. cv. Nipponbare) PIF family, in regulating seedling growth. OsPIL15 encodes a basic helix-loop-helix factor localized in the nucleus. OsPIL15-OX seedlings exhibit an exaggerated shorter above- ground part and undeveloped root system relative to wild-type seedlings, suggesting that OsPIL15 represses seedling growth in the dark. Microarray analysis combined with gene ontology analysis revealed that OsPIL15 represses a set of genes involved in auxin pathways and cell wall organization or biogenesis. Given the important roles of the auxin pathway and cell wall properties in controlling plant growth, we speculate that OsPIL15 represses seedling growth likely by regulating the auxin pathway and suppressing cell wall organization in etiolated rice seedlings. Additionally, exposure to red light or far-red light relieved growth retardation and promoted seedling elongation in the OsPIL15-OX lines, despite higher levels of OsPIL15 transcripts under red light and far-red light than in the dark. These results suggest that light regulation of OsPIL15 expres- sion is probably involved in photomorphogenesis in rice.
文摘Proteomic analysis was performed on seedlings after different light treatments. A total of (1 350±31) protein spots was separated and visualized on each silver nitrate-stained two-dimensional gel using protein samples prepared from light-grown or etiolated seedlings with or without 6-9 h light treatment. Twenty-five protein spots (encoded by 19 genes) that were significantly accumulated upon light treatment were identified using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry method. Functional proteomics indicated that these proteins involved mainly in chloroplast development, energy metabolism, cell cycle progression and membrane electron transport. For 18 of the protein-coding genes we identified through an internet search, the transcript levels of 17 genes matched roughly with their protein content in etiolated and green seedlings, suggesting that these genes were regulated by light mainly at the transcriptional level. Despite a very significant increase in the amount of proteins upon light treatment, similar RNA levels were found in dark-grown or green seedlings for the carbonic anhydrase gene At3g05100, indicating a possible post-transcriptional regulatory mechanism. Elucidation of light-induced protein accumulation will undoubtedly enhance our understanding of plant photomorphogenesis.
基金This work was supported by the National Science Foundation,by the Plant Biological Sciences Doctoral Dissertation Fellowship and Summer Fellowship from the University of Minnesota
文摘Ambient light has profound effects on early seedling de-etiolation through red and far-red light-absorbing phytochromes and blue and UV-A light-absorbing cryptochromes. Subsequent integration of various light signal trans- duction pathways leads to changes in gene expression and morphogenic responses. Here, we report the isolation of a new Arabidopsis light-signaling component, HYPOSENSITIVE TO LIGHT or HTL. Both htl-1 and htl-2 alleles displayed a long hypocotyl phenotype under red, far-red, and blue light, whereas overexpression of HTL caused a short hypocotyl pheno- type under similar light conditions. The mutants also showed other photomorphogenic defects such as elongated petioles, retarded cotyledon and leaf expansion, reduced accumulation of chlorophyll and anthocyanin pigments, and attenuated expression of light-responsive CHLOROPHYLL A/B BINDING PROTEIN 3 and CHALCONE SYNTHASE genes. HTL belongs to an alpha/beta fold protein family and is localized strongly in the nucleus and weakly in the cytosol. The expression of HTL was strongly induced by light of various wavelengths and this light induction was impaired in elongated hypocotyl 5. HY5 directly bound to both a C/G-box and a G-box in the HTL promoter but with a greater affinity toward the C/G-box. HTL, therefore, represents a new signaling step downstream of HY5 in phy- and cry-mediated de-etiolation responses.
基金supported by grants from the Korea Ocean Research & Development Institute (PE98474)by grants from BioGreen 21 Project funded by Rural Development Administration of Korea (20070401-034-028-009)
文摘Soluble invertase was purified from pea (Pisum sativum L.) by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid (SAC) and alkaline (SALK) invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively.