The stimulatory effect of lysophosphatidylcholine (lyso_PC) on ATP and ρ_nitrophenyl phosphate (PNPP) hydrolysis by the plasma membrane H +_ATPase from soybean (Glycine max (L.) Merr.) hypocotyls was studied. Re...The stimulatory effect of lysophosphatidylcholine (lyso_PC) on ATP and ρ_nitrophenyl phosphate (PNPP) hydrolysis by the plasma membrane H +_ATPase from soybean (Glycine max (L.) Merr.) hypocotyls was studied. Results showed that lyso_PC stimulated the hydrolysis of ATP; ATP hydrolysis was enhanced dramatically when lyso_PC was within 0-0.03%, and increased slightly when lyso_PC was higher than 0.03%. At the concentration of 0.03%, lyso_PC stimulated ATP hydrolysis by 80.5%. Kinetics analysis showed that V max increased from 0.46 μmol P i·mg -1 protein·min -1 to 0.87 μmol P i·mg -1 protein·min -1 while K m increased from 0.88 mmol/L to 1.15 mmol/L under lyso_PC treatment. The optimum pH of ATP hydrolysis was shifted from 6.5 to 7.0 . Moreover, it was found lyso_PC enhanced the inhibition of ATP hydrolysis by hydroxylamine. In the presence of 200 mmol/L hydroxylamine, ATP hydrolysis was inhibited by 74.4%, while it was inhibited by 84.4% when treated with lyso_PC. However, PNPP hydrolysis and the inhibitory effect of vanadate were not affected by lyso_PC. The above results indicated that the kinase domain might be an action site or regulatory region of the C_terminal autoinhibitory domain in the plant plasma membrane H +_ATPase.展开更多
The plasma membrane vesicles were purified from soybean (Glycine max L.) hypocotyls by two_phase partitioning methods. The stimulatory effects of K + on the coupling between ATP hydrolysis and proton transport by th...The plasma membrane vesicles were purified from soybean (Glycine max L.) hypocotyls by two_phase partitioning methods. The stimulatory effects of K + on the coupling between ATP hydrolysis and proton transport by the plasma membrane H +_ATPase were studied. The results showed that the proton transport activity was increased by 850% in the presence of 100 mmol/L KCl, while ATP hydrolytic activity was only increased by 28.2%. Kinetic studies showed that K m of ATP hydrolysis decreased from 1.14 to 0.7 mmol/L, while V max of ATP hydrolysis increased from 285.7 to 344.8 nmol Pi·mg -1 protein·min -1 in the presence of KCl. Experiments showed that the optimum pH was 6.5 and 6.0 in the presence and absence of KCl, respectively. Further studies revealed that K + could promote the inhibitory effects of hydroxylamines and vanadates on the ATP hydrolytic activity. The above results suggested that K + could regulate the coupling between ATP hydrolysis and proton transport of the plasma membrane H +_ATPase through modulating the structure and function of the kinase and phosphatase domains of the plasma membrane H +_ATPase.展开更多
Gravitropic curvature growth of Arabidopsis hypocotyls mainly occurred in the rapid growing Elongation Zone(EZI),not in the slow-growing Elongation Zone(EZII).By examining reorientation of Microtubules(MT)and phenotyp...Gravitropic curvature growth of Arabidopsis hypocotyls mainly occurred in the rapid growing Elongation Zone(EZI),not in the slow-growing Elongation Zone(EZII).By examining reorientation of Microtubules(MT)and phenotype of the cell wall in the EZI and the EZII of Arabidopsis hypocotyls under normal gravitational condition,it is found that MTs in the rapid growing epidermal cells were mainly in the transverse direction,while those in the non-growing epidermal cells were in the longitudinal directions.However,this difference in cortical MT arrays between the EZI and EZII cells disappeared when the seedlings were exposed to the simulated microgravity condition on a horizontal clinostat.Field emission scanning electron microscopy revealed that the surface texture of epidermal cells,like the direction of the MT,in the EZI and the EZII also became similar when exposed to the simulated microgravity condition.This result indicated that simulate microgravity could modify the potential differentiation between the EZI and the EZII by affecting the orientation of cortical MT in the epidermal cells.展开更多
Stomata are epidermal pores that are essential for water evaporation and gas exchange in plants.Stomatal development is orchestrated by intrinsic developmental programs,hormonal controls,and environmental cues.The ste...Stomata are epidermal pores that are essential for water evaporation and gas exchange in plants.Stomatal development is orchestrated by intrinsic developmental programs,hormonal controls,and environmental cues.The steroid hormone brassinosteroid(BR)inhibits stomatal lineage progression by regulating BIN2 and BSL proteins in leaves.Notably,BR is known to promote stomatal development in hypocotyls as opposed to leaves;however,its molecular mechanism remains elusive.Here,we show that BR signaling has a dual regulatory role in controlling stomatal development in Arabidopsis hypocotyls.We found that brassinolide(BL;the most active BR)regulates stomatal development differently in a concentration-dependent manner.At low and moderate concentrations,BL promoted stomatal formation by upregulating the expression of SPEECHLESS(SPCH)and its target genes independently of BIN2 regulation.In contrast,high concentrations of BL and bikinin,which is a specific inhibitor of BIN2 and its homologs,significantly reduced stomatal formation.Genetic analyses revealed that BIN2 regulates stomatal development in hypocotyls through molecular mechanisms distinct from the regulatory mechanism of the cotyledons.In hypocotyls,BIN2 promoted stomatal development by inactivating BZR1,which suppresses the expression of SPCH and its target genes.Taken together,our results suggest that BR precisely coordinates the stomatal development of hypocotyls using an antagonistic control of SPCH expression via BZR1-dependent and BZR1-independent transcriptional regulation.展开更多
Ethylene signaling is a complex pathway that has been intensively analyzed partly due to its importance to the manifestation of horticultural phenomena, including fruit ripening and tissue senescence. In order to furt...Ethylene signaling is a complex pathway that has been intensively analyzed partly due to its importance to the manifestation of horticultural phenomena, including fruit ripening and tissue senescence. In order to further our un- derstanding of how this pathway is regulated, a screen for Arabidopsis mutants with increased ethylene response was conducted. From this, a mutant was identified as having a dark-grown hypocotyl that is indistinguishable from Col-0 wt in the presence of the ethylene perception inhibitor AgNO3, yet has extreme responsiveness to even low levels of ethylene. Map-based cloning of the mutation revealed a T-DNA insertion in the coding sequence of the receptor-like kinase FERONIA, which is required for normal pollen tube reception and cell elongation in a currently unknown capacity. In contrast to a previous report, analysis of our feronia knockout mutant shows it also has altered responsiveness to brassinosteroids, with etiolated fer-2 seedlings being partially brassinosteroid insensitive with regard to promotion of hypocotyl elonga- tion. Our results indicate that FERONIA-dependent brassinosteroid response serves to antagonize the effect of ethylene on hypocotyl growth of etiolated seedlings, with loss of proper brassinosteroid signaling disrupting this balance and leading to a greater impact of ethylene on hypocotyl shortening.展开更多
Pepper(Capsicum annuum L.)is a typical self-pollinating crop with obvious heterosis in hybrids.Consequently,the use of morphological markers during the pepper seedling stage is crucial for pepper breeding.The color of...Pepper(Capsicum annuum L.)is a typical self-pollinating crop with obvious heterosis in hybrids.Consequently,the use of morphological markers during the pepper seedling stage is crucial for pepper breeding.The color of hypocotyl is widely used as a phenotypic marker in crossing studies of pepper.Pepper accessions generally have purple hypocotyls,which are mainly due to the anthocyanin accumulation in seedlings,and green hypocotyls are rarely observed in pepper.Here we reported the characterization of a green hypocotyl mutant of pepper,Cha1,which was identified from a pepper ethyl methanesulfonate(EMS)mutant library.Fine mapping revealed that the causal gene,CaTTG1,belonging to the WD40 repeat family,controlled the green hypocotyl phenotype of the mutant.Virus-induced gene silencing(VIGS)confirmed that CaTTG1 regulated anthocyanin accumulation.RNA-seq data showed that expression of structural genes CaDFR,CaANS,and CaUF3GT in the anthocyanin biosynthetic pathway was significantly decreased in Cha1 compared to the wild type.Yeast two-hybrid(Y2H)experiments also confirmed that CaTTG1 activated the synthesis of anthocyanin structural genes by forming a MBW complex with CaAN1 and CaGL3.In summary,this study provided a green hypocotyl mutant of pepper,and the Kompetitive Allele Specific PCR(KASP)marker developed based on the mutation site of the underlying gene would be helpful for pepper breeding.展开更多
Melon is a globally cultivated horticultural crop with a predominantly hybrid commercial seed market in China.Seedling morphology,particularly hypocotyl color,is a valuable trait for rapid F1 hybrid seed purity assess...Melon is a globally cultivated horticultural crop with a predominantly hybrid commercial seed market in China.Seedling morphology,particularly hypocotyl color,is a valuable trait for rapid F1 hybrid seed purity assessment.While green hypocotyls are common,white hypocotyls are rare in melon germplasm.This study identified a mutant with white hypocotyl but green leaves from the heavy ion beam mutant library.Genetic analysis revealed that a single recessive gene controlled the white hypocotyl,designated CmGhc1.A single-base deletion in the fifth exon of CmGhc1 led to a truncated CmGhc1 lacking the HTH-MYB DNA binding domain,likely affecting its transcriptional activity.CmGhc1 was localized in the nucleus,and yeast two-hybrid analysis and a dual-LUC assay demonstrated it as a transcription repressor.Furthermore,a KASP marker(hc1)was developed and verified as a functional marker for breeding white hypocotyl germplasms in melon.RNA-seq data revealed that CmGhc1 significantly affected the transcription of genes related to chlorophyll metabolism and photosynthesis in hypocotyl.In summary,these findings contribute to our understanding of chloroplast biogenesis and provide a valuable tool for melon breeding.展开更多
Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a...Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a comparative observation on the early hypocotyl development between two cultivars,‘SZQ’belonging to pak-choi(B.campestris ssp.chinensis var.communis)with longer hypocotyls,and‘WTC’belonging to Tacai(B.campestris L.ssp.chinensis var.rosularis)with shortter hypocotyls,and found that the difference in auxin biosynthesis might contribute to the varied hypocotyl phenotype between these two cultivars.By applying GWAS analysis using a total of 226 B.campestris accessions,we identified that the AT-Hook motif nuclear localized(AHL)gene BcAHL24-MF1 contributed to the natural variation in hypocotyl length.Functional variation of BcAHL24-MF1 was attributed to four haplotypes featuring four SNPs within the promoter region,of which Hap I accumulated more transcripts with shorter hypocotyls.Constitutive overexpression of BcAHL24-MF1 in B.campestris caused decreased hypocotyl length under light circumstances and even constant darkness,as BcAHL24-MF1 repressed the PIFmediated transcriptional activation of auxin biosynthesis genes BcYUC6-MF2 and BcYUC8-LF.Our research uncovered the important role of BcAHL24-MF1 in regulating light-triggered inhibition of hypocotyl elongation,therefore presenting a valuable genetic target for crop breeding.展开更多
The germination capacity of Ailanthus altissima seeds improved after the seeds were soaked with different concentrations of natural brassinolide （NBR）. The germination rate and germination energy of the seeds increa...The germination capacity of Ailanthus altissima seeds improved after the seeds were soaked with different concentrations of natural brassinolide （NBR）. The germination rate and germination energy of the seeds increased by 17.6% and 18.8%, and the mean germination speed （i.e., germination time） of the seeds was shortened by 1.4 d under the optimal concentration （0.4 mg·L^-1） treatment, compared with the control. After hypocotyls of A.altissima were treated with NBR, the elongation of the hypocotyls increased. Among different concentrations of the NBR, 0.4 mg·L^-1 NBR appeared to be the optimal concentration for the elongation of A.altissima hypocotyls.展开更多
[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by deal...[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by dealing with Apocynum seeds. Its cotyledons, hypocotyls and shoot tips were cultured on the media containing different concentrations of hormones. Finally, the influence of different hormone combinations on differentiation of cotyledons and hypocotyls, rapid multiplication of shoot tips, rapid multiplication of regenerated shoots, and rooting of test-tube plantlets was com- pared. [Result] MS＋2.0 mg/L BA＋0.03 mg/L NAA and MS＋0.07 mg/L NAA were the optimum medium for inducing regenerated buds from cotyledons and hypocotyls re- spectively; MS＋2.0 mg/L BA＋0.02 mg/L NAA was the best medium for rapid multi- plication of shoot tips; MS＋1.9 mg/L BA＋I.7 mg/L NAA was the best medium for rapid multiplication of regenerated buds： and 1/2MS＋0.6 mg/L NAA was the best medium for inducing roots. [Conclusion] The optimum hormone combination was de- termined for Apocynum rapid multiplication by tissue culture, which provides technical support on Apocynum industrialized cultivation.展开更多
[ Objective] This study aimed to optimize the in vitro regeneration system of melon. [ Method] Melon variety Nanxiang91023 was selected as experi- mental materials, with the cotyledons and hypocotyls as explants, diff...[ Objective] This study aimed to optimize the in vitro regeneration system of melon. [ Method] Melon variety Nanxiang91023 was selected as experi- mental materials, with the cotyledons and hypocotyls as explants, different types and concentrations of growth regulators were supplemented at different stages of tis- sue culture, to explore the simple and effective medium formula for regeneration of melon. [ Result] MS ＋ 1.5 mg/L of 6-BA ＋0.2 mg/L of LAA was the optimal medium for induction and proliferation of callus; MS ＋ 1.0 mg/L of 6-BA ＋ 0. 2 mg/L of 2, 4-D was the optimal medium for differentiation of adventitious buds; MS ＋ 1.0 mg/L of ZT ＋ 0.2 mg/L of LAA was the optimal medium for rooting of seedlings. On the basis of above conditions, melon seedlings had high roofing rate and strong roots. [ Conclusion] This study provided a guarantee for the further genetic transformation of improved melon varieties.展开更多
Using the hypocotyl and cotyledon explants of Brassica napus L. cuhivar Qingza No. 5 as receptors, hormone combinations in bud differentiation medi- um, bud growth medium and rooting medium were optimized to establish...Using the hypocotyl and cotyledon explants of Brassica napus L. cuhivar Qingza No. 5 as receptors, hormone combinations in bud differentiation medi- um, bud growth medium and rooting medium were optimized to establish an efficient plantlet regeneration system of B. napus cuhivar Qingza No. 5. The results showed that the highest differentiation efficiency of hypocotyls of B. napus cuhivar Qingza No. 5 reached about 90%, which was three times that of cotyledons. The appropriate differentiation medium was MSB ＋ 5 mg/L thidiazuron （TDZ） ＋7.5 mg/L AgNO3 ＋ 0.1 mg/L NAA ＋ 2 mg/L proline （L-pro） ＋ 250 mg/L casein acid hydrolysate （CH） ＋ 3% sucrose; the appropriate growth medium was 1/2 MSB ＋ 1 mg/L IBA ＋ 2 mg/L L-pro ＋ 250 mg/L CH ＋ 1.5% sucrose; the ap- propriate rooting medium was 1/2 MSB ＋ 0.2 mg/L IAA ＋ 1.5% sucrose. On this basis, a binary expression vector harboring insect resistance gene B12 was constructed and introduced into B. napus hypocotyls by Agrobacterium-mediated transformation. Positive plants were screened using hygromycin and carbenicillin. Transgenic plants were verified by PCR and GUS histochemical staining. The results showed that insect resistance gene B12 was successfully integrated into the nu- clear genome of B. napus plants and could be expressed normally. Leaves of transgenic plants with high expression levels were collected for indoor inoculation test with Plutella xylotella larvae to evaluate insect resistance of transgenic plants.展开更多
This study aimed to induce callus from three sunflower （Helianthus annulus L.） hybrids, namely Anna, Alhaja and Kuds, and to evaluate their callus for salt stress tolerance. Cotyledons and hypocotyl were taken from ...This study aimed to induce callus from three sunflower （Helianthus annulus L.） hybrids, namely Anna, Alhaja and Kuds, and to evaluate their callus for salt stress tolerance. Cotyledons and hypocotyl were taken from seedling of these hybrids and cultured on MS media contained 2,4-D （0,0, 0.5, 1.0, 1.5 and 2.0 mg/L） and kinetin （0.0, 0.5, and 1.0 mg/L）. The cultures incubated at 25 ＋ I^C under light condition （1,000 Lux） for 16 h/day. After 6 weeks observations were taken on the response of cotyledons and hypocotyl to callus induction. The induced callus were cultured on the same MS media that contained appropriate concentrations of 2,4-D and kinetin for callus induction as well as contained various concentration of sodium chloride NaCI （0.0, 0.05, 0.1, 0.15 and 0.2%）. After six weeks callus fresh and dry weights, proline and total carbohydrates concentrations were measured. The results showed significant differences among the hybrids, explants, 2,4-D and kinetin concentration and significant interaction between them in their percentage response for callus induction. The results also revealed that fresh and dry weights were significantly reduced with increased NaCI concentration in the medium, hybrids showed significant differences in their response to salt stress. Proline and total carbohydrate concentration increased in callus as NaCI increased in the media. Significant interaction was showed between hybrids and NaCI concentration in these parameters.展开更多
In this study, based on the simulated discharge results of chemical disinfectants, hypocotyl germination concentration gradient pre-test and concentration gradient determination experiment were set up respectively. La...In this study, based on the simulated discharge results of chemical disinfectants, hypocotyl germination concentration gradient pre-test and concentration gradient determination experiment were set up respectively. Laboratory cultivation was conducted to compare and analyze the root germination and germination indexes, three mangrove hypocotyls of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. Rhynchopetalas’ efficiency of cumulative root germination, cumulative germination and the cumulative expansion of the second pair of leaves, one-way analysis of variance was used to obtain the tolerance threshold of three mangrove hypocotyls to strong chlorin disinfectant. The study determined that the by-products of strong chlorin disinfectant, the toxic threshold concentrations of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. rhynchopetala are close to 0.55 mg/L, 0.55 mg/L and 0.25 mg/L, respectively. This concentration range is lower than the average concentration of 1.183 mg/L of active chlorine emitted from strong chlorine concentrate during pond clearing in high-level shrimp ponds, indicating that transient emissions of strong chlorine concentrate during pond clearing can have a toxic effect on mangrove plants. The strength of tolerance of the embryonic axes of the three mangrove species to effective chlorine contamination was, Ceriopstagal C.B. Rob. stronger than Bruguiera sexangula var. rhynchopetala, and Kandelia candel (Linn.) Druce is the weakest.展开更多
An efficient plant regeneration protocol was established for castor bean （Ricinus communis L.）, in which 0.3 mg L-1 thidiazuron （TDZ） induced shoot clusters and increased the number of adventitious shoots from hyp...An efficient plant regeneration protocol was established for castor bean （Ricinus communis L.）, in which 0.3 mg L-1 thidiazuron （TDZ） induced shoot clusters and increased the number of adventitious shoots from hypocotyl tissue. Our results showed that treatment under dark conditions significantly promoted the average number of shoots per explant to 37.36±4.54 （with a 6-d treatment）. Modified 1/2 Murashige and Skoog （MS） basal medium supplemented with 440 mg L-1 Ca2＋, 0.2 mg L-1 gibberellic acid and 0.1 mg L-1 TDZ significantly increased shoot elongation rates and lowered vitrification rates. Further- more, 1/2 MS media supplemented with 0.2 mg L-11-naphthaleneacetic acid induced a higher rooting rate compared with other culture conditions.展开更多
Ambient temperature induces the hypocotyl elongation of seedling,called as thermomorphogenesis.It has been reported that the bHLH transcriptional factor PIF7 acts as the critical component to modulate plant thermomorp...Ambient temperature induces the hypocotyl elongation of seedling,called as thermomorphogenesis.It has been reported that the bHLH transcriptional factor PIF7 acts as the critical component to modulate plant thermomorphogenesis,but the underlying mechanism remains elusive.The phytohormone abscisic acid(ABA)suppresses the hypocotyl elongation under high temperature(HT)stress.As the ABI5 binding protein,AFP2 acts as the negative factor to control ABA signaling.In this study,we first identified AFP2 as the interaction protein of PIF7 in vitro and in vivo.Phenotype analysis revealed that overexpressing AFP2 reduced the hypocotyl elongation,while loss-of-function afp2 mutant showed longer hypocotyl under HT.Consistently,overexpressing AFP2 impaired the transactivation effect of PIF7 on auxin biosynthesis related genes YUC8 and IAA19,which possibly resulted into the shorter hypocotyl in the transgenic line overexpressing AFP2 or co-overexpressing AFP2 and PIF7.Thus,these data suggest that AFP2 suppressed PIF7 activity to suppress hypocotyl elongation.Furthermore,we found that HT gradually induced the degradation of AFP2 that possibly released the inhibitory effect of AFP2 on PIF7,thus induced hypocotyl elongation under HT.Taken together,our result reveals the novel function of AFP2 in coordinating thermomorphogenesis through sophistically modulating PIF7 activity.展开更多
Seeds of soybeans and mung beans were soaked into five different concentrations （0, 0.05, 0.10, 0.20, 0.30 mg/L） of natural brassinolide （ NBR ） solution. According to the results, natural brassinolide treatment c...Seeds of soybeans and mung beans were soaked into five different concentrations （0, 0.05, 0.10, 0.20, 0.30 mg/L） of natural brassinolide （ NBR ） solution. According to the results, natural brassinolide treatment could improve seed germination rate and hypocotyl-radicle ratio of soybeans and mung beans and promote the growth of sprouts. To be specific, seed germination rate of soybeans reached the highest in 0.05 mg/L natural brassinolide treatment, which was im- proved by 25.0 percentage points compared with the control group, and the sprout length was improved by 4.33 em; 0.30 mg/L natural brassinolide exhibited the most significant promotion effect on seed germination and sprout growth of mung beans, seed germination rate was improved by 18.3 percentage points and sprout length was improved by 7.29 cm; hypocotyl-radiele ratio of soybean and mung bean sprouts reached the highest （2.96, 1.43 ） in 0.05 mg/L natural brassinolide treatment.展开更多
文摘The stimulatory effect of lysophosphatidylcholine (lyso_PC) on ATP and ρ_nitrophenyl phosphate (PNPP) hydrolysis by the plasma membrane H +_ATPase from soybean (Glycine max (L.) Merr.) hypocotyls was studied. Results showed that lyso_PC stimulated the hydrolysis of ATP; ATP hydrolysis was enhanced dramatically when lyso_PC was within 0-0.03%, and increased slightly when lyso_PC was higher than 0.03%. At the concentration of 0.03%, lyso_PC stimulated ATP hydrolysis by 80.5%. Kinetics analysis showed that V max increased from 0.46 μmol P i·mg -1 protein·min -1 to 0.87 μmol P i·mg -1 protein·min -1 while K m increased from 0.88 mmol/L to 1.15 mmol/L under lyso_PC treatment. The optimum pH of ATP hydrolysis was shifted from 6.5 to 7.0 . Moreover, it was found lyso_PC enhanced the inhibition of ATP hydrolysis by hydroxylamine. In the presence of 200 mmol/L hydroxylamine, ATP hydrolysis was inhibited by 74.4%, while it was inhibited by 84.4% when treated with lyso_PC. However, PNPP hydrolysis and the inhibitory effect of vanadate were not affected by lyso_PC. The above results indicated that the kinase domain might be an action site or regulatory region of the C_terminal autoinhibitory domain in the plant plasma membrane H +_ATPase.
文摘The plasma membrane vesicles were purified from soybean (Glycine max L.) hypocotyls by two_phase partitioning methods. The stimulatory effects of K + on the coupling between ATP hydrolysis and proton transport by the plasma membrane H +_ATPase were studied. The results showed that the proton transport activity was increased by 850% in the presence of 100 mmol/L KCl, while ATP hydrolytic activity was only increased by 28.2%. Kinetic studies showed that K m of ATP hydrolysis decreased from 1.14 to 0.7 mmol/L, while V max of ATP hydrolysis increased from 285.7 to 344.8 nmol Pi·mg -1 protein·min -1 in the presence of KCl. Experiments showed that the optimum pH was 6.5 and 6.0 in the presence and absence of KCl, respectively. Further studies revealed that K + could promote the inhibitory effects of hydroxylamines and vanadates on the ATP hydrolytic activity. The above results suggested that K + could regulate the coupling between ATP hydrolysis and proton transport of the plasma membrane H +_ATPase through modulating the structure and function of the kinase and phosphatase domains of the plasma membrane H +_ATPase.
基金Supported by the China Manned Space Flight Technology Project TG-2the National Natural Science Foundation of China(31670864)+2 种基金the National Natural Fund Joint Fund Project(U1738106)the Strategic Pioneer Projects of CAS(XDA15013900)the National Science Foundation for Young Scientists of China(31500687)
文摘Gravitropic curvature growth of Arabidopsis hypocotyls mainly occurred in the rapid growing Elongation Zone(EZI),not in the slow-growing Elongation Zone(EZII).By examining reorientation of Microtubules(MT)and phenotype of the cell wall in the EZI and the EZII of Arabidopsis hypocotyls under normal gravitational condition,it is found that MTs in the rapid growing epidermal cells were mainly in the transverse direction,while those in the non-growing epidermal cells were in the longitudinal directions.However,this difference in cortical MT arrays between the EZI and EZII cells disappeared when the seedlings were exposed to the simulated microgravity condition on a horizontal clinostat.Field emission scanning electron microscopy revealed that the surface texture of epidermal cells,like the direction of the MT,in the EZI and the EZII also became similar when exposed to the simulated microgravity condition.This result indicated that simulate microgravity could modify the potential differentiation between the EZI and the EZII by affecting the orientation of cortical MT in the epidermal cells.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korean government(Ministry of Science and ICT or Ministry of Education)(2021R1A2C1006617 and RS-2024-00407469 to T.-W.K.)supported by the Korea Basic Science Institute(National Research Facilities and Equipment Center)grant funded by the Ministry of Education(2023R1A6C101A009 to T.-K.P.)the research fund of Hanyang University(HY-202200000003024 to T.-K.P.)。
文摘Stomata are epidermal pores that are essential for water evaporation and gas exchange in plants.Stomatal development is orchestrated by intrinsic developmental programs,hormonal controls,and environmental cues.The steroid hormone brassinosteroid(BR)inhibits stomatal lineage progression by regulating BIN2 and BSL proteins in leaves.Notably,BR is known to promote stomatal development in hypocotyls as opposed to leaves;however,its molecular mechanism remains elusive.Here,we show that BR signaling has a dual regulatory role in controlling stomatal development in Arabidopsis hypocotyls.We found that brassinolide(BL;the most active BR)regulates stomatal development differently in a concentration-dependent manner.At low and moderate concentrations,BL promoted stomatal formation by upregulating the expression of SPEECHLESS(SPCH)and its target genes independently of BIN2 regulation.In contrast,high concentrations of BL and bikinin,which is a specific inhibitor of BIN2 and its homologs,significantly reduced stomatal formation.Genetic analyses revealed that BIN2 regulates stomatal development in hypocotyls through molecular mechanisms distinct from the regulatory mechanism of the cotyledons.In hypocotyls,BIN2 promoted stomatal development by inactivating BZR1,which suppresses the expression of SPCH and its target genes.Taken together,our results suggest that BR precisely coordinates the stomatal development of hypocotyls using an antagonistic control of SPCH expression via BZR1-dependent and BZR1-independent transcriptional regulation.
文摘Ethylene signaling is a complex pathway that has been intensively analyzed partly due to its importance to the manifestation of horticultural phenomena, including fruit ripening and tissue senescence. In order to further our un- derstanding of how this pathway is regulated, a screen for Arabidopsis mutants with increased ethylene response was conducted. From this, a mutant was identified as having a dark-grown hypocotyl that is indistinguishable from Col-0 wt in the presence of the ethylene perception inhibitor AgNO3, yet has extreme responsiveness to even low levels of ethylene. Map-based cloning of the mutation revealed a T-DNA insertion in the coding sequence of the receptor-like kinase FERONIA, which is required for normal pollen tube reception and cell elongation in a currently unknown capacity. In contrast to a previous report, analysis of our feronia knockout mutant shows it also has altered responsiveness to brassinosteroids, with etiolated fer-2 seedlings being partially brassinosteroid insensitive with regard to promotion of hypocotyl elonga- tion. Our results indicate that FERONIA-dependent brassinosteroid response serves to antagonize the effect of ethylene on hypocotyl growth of etiolated seedlings, with loss of proper brassinosteroid signaling disrupting this balance and leading to a greater impact of ethylene on hypocotyl shortening.
基金supported by grants from the Special Funds for Construction of Innovative Provinces in Hunan Province(Grant No.2021NK1006)the Science and Technology Innovation Program of Hunan Province(Grant No.2021JC0007)+2 种基金China Agriculture Research System of MOF and MARA(Grant No.CARS-24-A-15)National Natural Science Foundation of China(Grant No.32130097)National Natural Science Foundation of China(Grant No.U19A2028)。
文摘Pepper(Capsicum annuum L.)is a typical self-pollinating crop with obvious heterosis in hybrids.Consequently,the use of morphological markers during the pepper seedling stage is crucial for pepper breeding.The color of hypocotyl is widely used as a phenotypic marker in crossing studies of pepper.Pepper accessions generally have purple hypocotyls,which are mainly due to the anthocyanin accumulation in seedlings,and green hypocotyls are rarely observed in pepper.Here we reported the characterization of a green hypocotyl mutant of pepper,Cha1,which was identified from a pepper ethyl methanesulfonate(EMS)mutant library.Fine mapping revealed that the causal gene,CaTTG1,belonging to the WD40 repeat family,controlled the green hypocotyl phenotype of the mutant.Virus-induced gene silencing(VIGS)confirmed that CaTTG1 regulated anthocyanin accumulation.RNA-seq data showed that expression of structural genes CaDFR,CaANS,and CaUF3GT in the anthocyanin biosynthetic pathway was significantly decreased in Cha1 compared to the wild type.Yeast two-hybrid(Y2H)experiments also confirmed that CaTTG1 activated the synthesis of anthocyanin structural genes by forming a MBW complex with CaAN1 and CaGL3.In summary,this study provided a green hypocotyl mutant of pepper,and the Kompetitive Allele Specific PCR(KASP)marker developed based on the mutation site of the underlying gene would be helpful for pepper breeding.
基金financially supported by the grants from the earmarked fund for Xinjiang Agriculture Research System,China(XJARS-06)the Key Research and Development Task Special Project of Xinjiang,China(2022B02002-3)+1 种基金the Tianshan Talent Training Program,China(2023TSYCLJ0015)the Key Research and Development Program of Xinjiang Uygur Autonomous Region,China(2023B02017).
文摘Melon is a globally cultivated horticultural crop with a predominantly hybrid commercial seed market in China.Seedling morphology,particularly hypocotyl color,is a valuable trait for rapid F1 hybrid seed purity assessment.While green hypocotyls are common,white hypocotyls are rare in melon germplasm.This study identified a mutant with white hypocotyl but green leaves from the heavy ion beam mutant library.Genetic analysis revealed that a single recessive gene controlled the white hypocotyl,designated CmGhc1.A single-base deletion in the fifth exon of CmGhc1 led to a truncated CmGhc1 lacking the HTH-MYB DNA binding domain,likely affecting its transcriptional activity.CmGhc1 was localized in the nucleus,and yeast two-hybrid analysis and a dual-LUC assay demonstrated it as a transcription repressor.Furthermore,a KASP marker(hc1)was developed and verified as a functional marker for breeding white hypocotyl germplasms in melon.RNA-seq data revealed that CmGhc1 significantly affected the transcription of genes related to chlorophyll metabolism and photosynthesis in hypocotyl.In summary,these findings contribute to our understanding of chloroplast biogenesis and provide a valuable tool for melon breeding.
基金supported by grants from the Key R and D Program of Zhejiang(Grant Nos.2022C02032 and 2022C02030)the SanNong JiuFang Science and Technology Cooperation Project of Zhejiang Province(Grant No.2023SNJF008)+1 种基金the Grand Science and Technology Special Project of Zhejiang Province(Grant No.2021C02065)the Science and Technology Plan Project of Jiaxing(Grant No.2023AZ11002).
文摘Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a comparative observation on the early hypocotyl development between two cultivars,‘SZQ’belonging to pak-choi(B.campestris ssp.chinensis var.communis)with longer hypocotyls,and‘WTC’belonging to Tacai(B.campestris L.ssp.chinensis var.rosularis)with shortter hypocotyls,and found that the difference in auxin biosynthesis might contribute to the varied hypocotyl phenotype between these two cultivars.By applying GWAS analysis using a total of 226 B.campestris accessions,we identified that the AT-Hook motif nuclear localized(AHL)gene BcAHL24-MF1 contributed to the natural variation in hypocotyl length.Functional variation of BcAHL24-MF1 was attributed to four haplotypes featuring four SNPs within the promoter region,of which Hap I accumulated more transcripts with shorter hypocotyls.Constitutive overexpression of BcAHL24-MF1 in B.campestris caused decreased hypocotyl length under light circumstances and even constant darkness,as BcAHL24-MF1 repressed the PIFmediated transcriptional activation of auxin biosynthesis genes BcYUC6-MF2 and BcYUC8-LF.Our research uncovered the important role of BcAHL24-MF1 in regulating light-triggered inhibition of hypocotyl elongation,therefore presenting a valuable genetic target for crop breeding.
文摘The germination capacity of Ailanthus altissima seeds improved after the seeds were soaked with different concentrations of natural brassinolide （NBR）. The germination rate and germination energy of the seeds increased by 17.6% and 18.8%, and the mean germination speed （i.e., germination time） of the seeds was shortened by 1.4 d under the optimal concentration （0.4 mg·L^-1） treatment, compared with the control. After hypocotyls of A.altissima were treated with NBR, the elongation of the hypocotyls increased. Among different concentrations of the NBR, 0.4 mg·L^-1 NBR appeared to be the optimal concentration for the elongation of A.altissima hypocotyls.
基金Supported by Science and Technology Development Program of Jilin Province(20110909)Youth Foud of Baicheng Normal University(200801)~~
文摘[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by dealing with Apocynum seeds. Its cotyledons, hypocotyls and shoot tips were cultured on the media containing different concentrations of hormones. Finally, the influence of different hormone combinations on differentiation of cotyledons and hypocotyls, rapid multiplication of shoot tips, rapid multiplication of regenerated shoots, and rooting of test-tube plantlets was com- pared. [Result] MS＋2.0 mg/L BA＋0.03 mg/L NAA and MS＋0.07 mg/L NAA were the optimum medium for inducing regenerated buds from cotyledons and hypocotyls re- spectively; MS＋2.0 mg/L BA＋0.02 mg/L NAA was the best medium for rapid multi- plication of shoot tips; MS＋1.9 mg/L BA＋I.7 mg/L NAA was the best medium for rapid multiplication of regenerated buds： and 1/2MS＋0.6 mg/L NAA was the best medium for inducing roots. [Conclusion] The optimum hormone combination was de- termined for Apocynum rapid multiplication by tissue culture, which provides technical support on Apocynum industrialized cultivation.
基金Supported by the Foundation of Hunan Provincial Department of Science and Technology(2010FJ3103)
文摘[ Objective] This study aimed to optimize the in vitro regeneration system of melon. [ Method] Melon variety Nanxiang91023 was selected as experi- mental materials, with the cotyledons and hypocotyls as explants, different types and concentrations of growth regulators were supplemented at different stages of tis- sue culture, to explore the simple and effective medium formula for regeneration of melon. [ Result] MS ＋ 1.5 mg/L of 6-BA ＋0.2 mg/L of LAA was the optimal medium for induction and proliferation of callus; MS ＋ 1.0 mg/L of 6-BA ＋ 0. 2 mg/L of 2, 4-D was the optimal medium for differentiation of adventitious buds; MS ＋ 1.0 mg/L of ZT ＋ 0.2 mg/L of LAA was the optimal medium for rooting of seedlings. On the basis of above conditions, melon seedlings had high roofing rate and strong roots. [ Conclusion] This study provided a guarantee for the further genetic transformation of improved melon varieties.
基金Supported by National Natural Science Foundation of China(31301703)Agricultural Science and Technology Independent Innovation Fund of Jiangsu Province[CX(14)5068]
文摘Using the hypocotyl and cotyledon explants of Brassica napus L. cuhivar Qingza No. 5 as receptors, hormone combinations in bud differentiation medi- um, bud growth medium and rooting medium were optimized to establish an efficient plantlet regeneration system of B. napus cuhivar Qingza No. 5. The results showed that the highest differentiation efficiency of hypocotyls of B. napus cuhivar Qingza No. 5 reached about 90%, which was three times that of cotyledons. The appropriate differentiation medium was MSB ＋ 5 mg/L thidiazuron （TDZ） ＋7.5 mg/L AgNO3 ＋ 0.1 mg/L NAA ＋ 2 mg/L proline （L-pro） ＋ 250 mg/L casein acid hydrolysate （CH） ＋ 3% sucrose; the appropriate growth medium was 1/2 MSB ＋ 1 mg/L IBA ＋ 2 mg/L L-pro ＋ 250 mg/L CH ＋ 1.5% sucrose; the ap- propriate rooting medium was 1/2 MSB ＋ 0.2 mg/L IAA ＋ 1.5% sucrose. On this basis, a binary expression vector harboring insect resistance gene B12 was constructed and introduced into B. napus hypocotyls by Agrobacterium-mediated transformation. Positive plants were screened using hygromycin and carbenicillin. Transgenic plants were verified by PCR and GUS histochemical staining. The results showed that insect resistance gene B12 was successfully integrated into the nu- clear genome of B. napus plants and could be expressed normally. Leaves of transgenic plants with high expression levels were collected for indoor inoculation test with Plutella xylotella larvae to evaluate insect resistance of transgenic plants.
文摘This study aimed to induce callus from three sunflower （Helianthus annulus L.） hybrids, namely Anna, Alhaja and Kuds, and to evaluate their callus for salt stress tolerance. Cotyledons and hypocotyl were taken from seedling of these hybrids and cultured on MS media contained 2,4-D （0,0, 0.5, 1.0, 1.5 and 2.0 mg/L） and kinetin （0.0, 0.5, and 1.0 mg/L）. The cultures incubated at 25 ＋ I^C under light condition （1,000 Lux） for 16 h/day. After 6 weeks observations were taken on the response of cotyledons and hypocotyl to callus induction. The induced callus were cultured on the same MS media that contained appropriate concentrations of 2,4-D and kinetin for callus induction as well as contained various concentration of sodium chloride NaCI （0.0, 0.05, 0.1, 0.15 and 0.2%）. After six weeks callus fresh and dry weights, proline and total carbohydrates concentrations were measured. The results showed significant differences among the hybrids, explants, 2,4-D and kinetin concentration and significant interaction between them in their percentage response for callus induction. The results also revealed that fresh and dry weights were significantly reduced with increased NaCI concentration in the medium, hybrids showed significant differences in their response to salt stress. Proline and total carbohydrate concentration increased in callus as NaCI increased in the media. Significant interaction was showed between hybrids and NaCI concentration in these parameters.
文摘In this study, based on the simulated discharge results of chemical disinfectants, hypocotyl germination concentration gradient pre-test and concentration gradient determination experiment were set up respectively. Laboratory cultivation was conducted to compare and analyze the root germination and germination indexes, three mangrove hypocotyls of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. Rhynchopetalas’ efficiency of cumulative root germination, cumulative germination and the cumulative expansion of the second pair of leaves, one-way analysis of variance was used to obtain the tolerance threshold of three mangrove hypocotyls to strong chlorin disinfectant. The study determined that the by-products of strong chlorin disinfectant, the toxic threshold concentrations of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. rhynchopetala are close to 0.55 mg/L, 0.55 mg/L and 0.25 mg/L, respectively. This concentration range is lower than the average concentration of 1.183 mg/L of active chlorine emitted from strong chlorine concentrate during pond clearing in high-level shrimp ponds, indicating that transient emissions of strong chlorine concentrate during pond clearing can have a toxic effect on mangrove plants. The strength of tolerance of the embryonic axes of the three mangrove species to effective chlorine contamination was, Ceriopstagal C.B. Rob. stronger than Bruguiera sexangula var. rhynchopetala, and Kandelia candel (Linn.) Druce is the weakest.
基金supported by the National Natural Science Foundation of China (31260336 and 31460353)
文摘An efficient plant regeneration protocol was established for castor bean （Ricinus communis L.）, in which 0.3 mg L-1 thidiazuron （TDZ） induced shoot clusters and increased the number of adventitious shoots from hypocotyl tissue. Our results showed that treatment under dark conditions significantly promoted the average number of shoots per explant to 37.36±4.54 （with a 6-d treatment）. Modified 1/2 Murashige and Skoog （MS） basal medium supplemented with 440 mg L-1 Ca2＋, 0.2 mg L-1 gibberellic acid and 0.1 mg L-1 TDZ significantly increased shoot elongation rates and lowered vitrification rates. Further- more, 1/2 MS media supplemented with 0.2 mg L-11-naphthaleneacetic acid induced a higher rooting rate compared with other culture conditions.
基金funded by the National Natural Science Foundation of China(Grant No.31970289).
文摘Ambient temperature induces the hypocotyl elongation of seedling,called as thermomorphogenesis.It has been reported that the bHLH transcriptional factor PIF7 acts as the critical component to modulate plant thermomorphogenesis,but the underlying mechanism remains elusive.The phytohormone abscisic acid(ABA)suppresses the hypocotyl elongation under high temperature(HT)stress.As the ABI5 binding protein,AFP2 acts as the negative factor to control ABA signaling.In this study,we first identified AFP2 as the interaction protein of PIF7 in vitro and in vivo.Phenotype analysis revealed that overexpressing AFP2 reduced the hypocotyl elongation,while loss-of-function afp2 mutant showed longer hypocotyl under HT.Consistently,overexpressing AFP2 impaired the transactivation effect of PIF7 on auxin biosynthesis related genes YUC8 and IAA19,which possibly resulted into the shorter hypocotyl in the transgenic line overexpressing AFP2 or co-overexpressing AFP2 and PIF7.Thus,these data suggest that AFP2 suppressed PIF7 activity to suppress hypocotyl elongation.Furthermore,we found that HT gradually induced the degradation of AFP2 that possibly released the inhibitory effect of AFP2 on PIF7,thus induced hypocotyl elongation under HT.Taken together,our result reveals the novel function of AFP2 in coordinating thermomorphogenesis through sophistically modulating PIF7 activity.
基金Supported by Key Joint Foundation for Fostering Talents of NSFC-Henan Province(U1204307)Key Project of Science and Technology of Henan Province(102102110155,142102110173,152102210334)Cultivation Fund of Luoyang Normal University(2013-PYJJ-001,10000993)
文摘Seeds of soybeans and mung beans were soaked into five different concentrations （0, 0.05, 0.10, 0.20, 0.30 mg/L） of natural brassinolide （ NBR ） solution. According to the results, natural brassinolide treatment could improve seed germination rate and hypocotyl-radicle ratio of soybeans and mung beans and promote the growth of sprouts. To be specific, seed germination rate of soybeans reached the highest in 0.05 mg/L natural brassinolide treatment, which was im- proved by 25.0 percentage points compared with the control group, and the sprout length was improved by 4.33 em; 0.30 mg/L natural brassinolide exhibited the most significant promotion effect on seed germination and sprout growth of mung beans, seed germination rate was improved by 18.3 percentage points and sprout length was improved by 7.29 cm; hypocotyl-radiele ratio of soybean and mung bean sprouts reached the highest （2.96, 1.43 ） in 0.05 mg/L natural brassinolide treatment.
