Cortical Microtubule Reorientation and Its Relation to Cell Surface Texture of Epidermal Cells of Arabidopsis Thaliana Hypocotyls under Simulated Microgravity Conditions

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.

The Toxic Effects of Strong Chlorin Disinfectant on Mangroves and Emission Thresholds

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.

FERONIA Is a Key Modulator of Brassinosteroid and Ethylene Responsiveness in Arabidopsis Hypocotyls

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.

The photosensory function of Zmphot1 differs from that of Atphot1 due to the C-terminus of Zmphot1 during phototropic response

The role of phot1 in triggering hypocotyl phototropism and optimizing growth orientation has been wellcharacterized in Arabidopsis, whereas the role of Zmphot1 in maize remains largely unclear. Here, we show that Zmphot1 is involved in blue light-induced phototropism. Compared with Atphot1, Zmphot1exhibited a weaker phototropic response to very low-fluence rates of blue light(< 0.01 μmol m-2s-1),but stronger phototropic response to high-fluence rates of blue light(> 10 μmol m-2s-1) than Atphot1. Notably, blue light exposure induced Zmphot1-green fluorescent protein(GFP), but not Atphot1-GFP, to form the aggregates in the cytoplasm of Nicotiana benthamiana cells. Furthermore, by generating the chimeric phot1 proteins, we found that the serine-threonine kinase(STK) domain at the C-terminus is responsible for a more volatile membrane association of Zmphot1. Consistently, the chimeric phot1 protein fusing the STK domain of Zmphot1 with other domains of Atphot1 responded similarly as Zmphot1 to both low and high fluence rates of blue light. Interestingly, although both Zmphot1 and Atphot1 interact with AtNPH3, Zmphot1 induced weaker dephosphorylation of NONPHOTOTROPIC HYPOCOTYL 3(NPH3) than Atphot1. Together, our findings indicate that Zmphot1 and Atphot1 exhibit different photosensory function during phototropic response and that the STK domain may play a key role in determining their properties.

Comparative Evaluation of Humic Substances: Effect at Cell Level and Chlorophyll Retention during Accelerated Senescence

The influence of humic substances (HS) formulations derived from sedimentary and compost sources was studied on plant growth at cell level and chlorophyll retention during accelerated senescence of leaf tissue. The direct effect of HS formulations was studied on cell expansion using cucumber and radish cotyledon expansion test. The cucumber hypocotyl elongation test was used to study the effect on cell elongation. Chlorophyll pigment retention in excised leaf tissue incubated in dark with high temperature was assessed to study the effect on leaf senescence. Explant tissues were incubated directly in the solutions of the formulations at the concentration recommended for foliar application to the crop plants. HS formulations showed significant variations in their direct bio-stimulatory effects. Formulations derived from compost sources were found superior in terms of inducing a direct stimulatory effect on cell expansion and cell elongation and in maintaining chlorophyll pigment retention during accelerated senescence. HS from sedimentary sources stimulated cell expansion and delayed chlorophyll degradation to a lesser extent compared to HS from compost. However, HS formulations derived from sedimentary sources used in this study were not effective in inducing cell elongation in the cucumber hypocotyl elongation test. The direct bio-stimulatory effect of HS formulations differed significantly between the formulations that were evaluated.

Effect of Natural Brassinolide on Germination of Ailanthus altissima Seeds

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.

Study on In vitro Regeneration System of Melon

[ 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.

Efficient Regeneration and Agrobacterium-mediated Transformation of Brassica napus Cultivar Qingza No.5

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.

日本结缕草ZjHY5的克隆、亚细胞定位及转录自激活检测

转录因子LONG HYPOCOTYL 5(HY5)在光诱导基因表达中起中心调控因子的作用。为了探究日本结缕草(Zoysia japonica)中ZjHY5基因的亚细胞定位及自激活特性,本研究从日本结缕草(Zoysia japonica)中克隆得到ZjHY5基因。该基因开放阅读框为711 bp,编码236个氨基酸残基。ZjHY5蛋白无信号肽,进化树分析显示其与漆姑草(Sagina japonica)及苜蓿(Medicago sativa)中HY5蛋白的相似度最高。亚细胞定位结果显示,ZjHY5定位于细胞核中。通过构建酵母诱饵载体,并对其进行自激活检测,结果显示,ZjHY5基因无自激活活性,可用于后续试验。本试验为深入研究日本结缕草ZjHY5基因的功能奠定了基础。

AFP2 Coordinates the Activity of PIF7 for Thermomorphogenesis in Arabidopsis Seedlings

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.

有机氯农药DDT对秋茄萌生的影响

有机氯农药ＤＤＴ对秋茄萌生的影响黄建斌，林鹏（生物学系环境科学研究中心）红树林是热带、亚热带河口海岸的木本植物群落￣［１］．它具有较强的生活力，对河口海区的污染物有一定净化能力，红树植物生长在有机机氯农药污染的水体中它将在生长上受到什么影响，尚未见过...

In vitro establishment of a highly effective method of castor bean (Ricinus communis L.) regeneration using shoot explants

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.

Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi

Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicus Cu zhi. Hypocotyl slices were cultured on Murashige and Skoog （MS） and B5 basal medium supplemented with varied concentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid （NAA） and indole-3-butyric acid （IBA） in combination with 6-benzylaminopurine （6-BA） and kinetin. The study showed that shoots could be directly regenerated from hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shoots regeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings. The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal medium supplemented with 1.5 mg L^-1 6-BA and 0.05 mg L^-1 NAA （63.64%）. The regenerated shoots were readily elongated on the same medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L^-1 IBA and 100 mg L^-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfully acclimatized. This regeneration system is applied for genetic transformation now.

Morphology and ontogeny of directly differentiating shoot buds and somatic embryos in Santalum album L.

Santalum album L.is a commercially important tree that yields essential oil of high medicinal value.Regeneration research through organogenesis and embryogenesis has been documented but no report depicts comparative ontogeny of directly differentiating shoot buds(SB)and somatic embryos(SE).In the present study aseptic seedling derived hypocotyl segments(HC)and hypocotyl+root junction(HC+R)were used to induce SBs and SEs,respectively.Ontogenic differences between the structures were confirmed using scanning electron microscopy and histological analysis.MS medium containing 6-benzyladenine or BA(2.5μM)produced highest number of direct SB,while MS+BA(7.5μM)proved suitable for higher frequency of SE differentiation.The differentiating structures attained growth when transferred to MS medium containing a combination of BA and anaphthalene acetic acid or silver nitrate(AgNO3).A combination of indole-3-butyric acid and silver nitrate(AgNO3)in half-strength woody plant medium and lesser osmotic concentration(2%sucrose),induced rhizogenesis.

Effects of Natural Brassinolide on Seed Germination and Hypocotyl Elongation of Beans

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.

Diversity and association analysis of important agricultural trait based on miniature inverted-repeat transposable element specific marker in Brassica napus L.

Miniature inverted-repeat transposable elements(MITEs)are a group of DNA transposable element(TE)which preferentially distributed with gene associated regions.Tens of MITEs families have been revealed in Brassica napus genome,they scatter across the genome with tens of thousands copies and produce polymorphisms both intra-and inter-species.Our previous studies revealed a Tourist-like MITE,Monkey King,associated with vernalization requirement of B.napus,however there are still few studies reveal MITE association with agricultural traits in B.napus.In the present study,80 polymorphic markers were developed from 55 MITEs,and used to evaluate genetic diversity in a panel of B.napus accessions consisting of 101 natural and 25 synthetic genotypes.Five agricultural traits,oil content,glucosinolate content,erucic acid content,weight of thousand seeds(WTS)and plant height,were investigated across 3-years field experiments,in addition,two traits,hypocotyl length and root length,were evaluated at the 4-leaf stage in the laboratory.Correlations between the MITE-based markers and seven traits were analyzed,finally,10 polymorphic markers produced by 6 pairs of MITE specific primers were revealed relatively high correlation with 5 traits.Two polymorphic markers were anchored with two candidate genes,BnaA02g13530D and BnaA08g20010D,respectively,which may contribute to glucosinolate content and WTS.This research may contribute to genetic improvement through utilization of MITE-induced polymorphisms in Brassica species.

Impacts of Light on Rooting of Mangrove Kandelia candel Propagules

The process of rooting of propagules is one of the critical aspects fordiscussing the ecological expansion of mangrove populations.By the experimentsof culture,we found out that this process was controlled by light in the viviparoushypocotyl of Kandelia candel.This discovery supplies a favourable access toexplain why the floating propagules of K.candel can remain alive for a long timeon the sea and be successfully dispersed by tides.

Effects of non-procedural factors in Brassica napus genetic transformation

Rapeseed (Brassica napus) is the second largest oil crop in the world. However, transformation efficiency of rapeseed still needs to be improved. To evaluate non-procedural factors (e.g. explants, section of explant, marker genes and number of exogenous genes) effects on transformation efficiency, 6-day-old hypocotyl explants from in vitro grown seedlings were co-cultivated with Agrobacterium strain GV3101 harboring a binary vector using optimized transformation procedure. Results showed that normal maturing variety ‘Zhongshuang 6 (ZS6)’ had the highest overall capacity to produce rooted shoots among 5 common varieties and 6 early maturing varieties, with green callus induction rate 81.45% and shoot regeneration rate 21.66%. Early maturing variety 14M645 has relatively high regeneration rate (4.69%) and one of the shortest growth periods (107.54 d). Data showed that choosing neomycin phosphotransferase II gene (NPTII) as selectable marker led to the best transformation rate (17.38%). Selecting upper hypocotyl segments near cotyledon as explant provided the higest transformation efficiency, with regeneration rate of 25.59% when using NPTII as selectable marker and 22.19% for Bar. B. napus transformed with single gene showed higher transformation frequency than vectors with multiple genes,highlighting difficulty of multiple gene transformation. This work helped to further improve genetic transformation of B. napus by optimizing factors that impact transformation efficiency,and it would ultimately improve research in transgenic B. napus varieties with commercial potential.

Candidate genes conferring ethylene-response in cultivated peanuts determined by BSA-seq and fine-mapping

Ethylene plays essential roles in plant growth,development and stress responses.The ethylene signaling pathway and molecular mechanism have been studied extensively in Arabidopsis and rice but limited in peanuts.Here,we established a sand-culture method to screen pingyangmycin mutagenized peanut lines based on their specific response to ethylene(“triple response”).An ethylene-insensitive mutant,inhibition of peanut hypocotyl elongation 1(iph1),was identified that showed reduced sensitivity to ethylene in both hypocotyl elongation and root growth.Through bulked segregant analysis sequencing,a major gene related to iph1,named AhIPH1,was preliminarily mapped at the chromosome Arahy.01,and further narrowed to a 450-kb genomic region through substitution mapping strategy.A total of 7014 genes were differentially expressed among the ACC treatment through RNA-seq analysis,of which only the Arahy.5BLU0Q gene in the candidate mapping interval was differentially expressed between WT and mutant iph1.Integrating sequence variations,functional annotation and transcriptome analysis revealed that a predicated gene,Arahy.5BLU0Q,encoding SNF1 protein kinase,may be the candidate gene for AhIPH1.This gene contained two single-nucleotide polymorphisms at promoter region and was more highly expressed in iph1 than WT.Our findings reveal a novel ethylene-responsive gene,which provides a theoretical foundation and new genetic resources for the mechanism of ethylene signaling in peanuts.

The DELLA-ABI4-HY5 module integrates light and gibberellin signals to regulate hypocotyl elongation

Plant growth is coordinately controlled by various environmental and hormonal signals,of which light and gibberellin(GA)signals are two critical factors with opposite effects on hypocotyl elongation.Although interactions between the light and GA signaling pathways have been studied extensively,the detailed regulatory mechanism of their direct crosstalk in hypocotyl elongation remains to be fully clarified.Previously,we reported that ABA INSENSITIVE 4(ABI4)controls hypocotyl elongation through its regulation of cellelongation-related genes,but whether it is also involved in GA signaling to promote hypocotyl elongation is unknown.In this study,we showthat promotion of hypocotyl elongation by GA is dependent on ABI4 activation.DELLAs interact directly with ABI4 and inhibit its DNA-binding activity.In turn,ABI4 combined with ELONGATED HYPOCOTYL 5(HY5),a key positive factor in light signaling,feedback regulates the expression of the GA2ox GA catabolism genes and thus modulates GA levels.Taken together,our results suggest that the DELLA-ABI4-HY5 module may serve as a molecular link that integrates GA and light signals to control hypocotyl elongation.