A bHLH transcription factor,CsSPT,regulates high-temperature resistance in cucumber

High-temperature stress threatens the growth and yield of crops. Basic helix-loop-helix(bHLH) transcription factors(TFs) have been shown to play important roles in regulating high-temperature resistance in plants. However, the bHLH TFs responsible for high-temperature tolerance in cucumbers have not been identified. We used transcriptome profiling to screen the high temperature-responsive candidate bHLH TFs in cucumber. Here, we found that the expression of 75 CsbHLH genes was altered under high-temperature stress. The expression of the CsSPT gene was induced by high temperatures in TT(Thermotolerant) cucumber plants. However, the Csspt mutant plants obtained by the CRISPR-Cas9 system showed severe thermosensitive symptoms, including wilted leaves with brown margins and reduced root density and cell activity.The Csspt mutant plants also exhibited elevated H_(2)O_(2) levels and down-regulated photosystem-related genes under normal conditions.Furthermore, there were high relative electrolytic leakage(REC), malondialdehyde(MDA), glutathione(GSH), and superoxide radical(O_(2)^(·-)) levels in the Csspt mutant plants, with decreased Proline content after the high-temperature treatment. Transcriptome analysis showed that the photosystem and chloroplast activities in Csspt mutant plants were extremely disrupted by the high-temperature stress compared with wildtype(WT) plants. Moreover, the plant hormone signal transduction, as well as MAPK and calcium signaling pathways were activated in Csspt mutant plants under high-temperature stress. The HSF and HSP family genes shared the same upregulated expression patterns in Csspt and WT plants under high-temperature conditions. However, most bHLH, NAC, and bZIP family genes were significantly down-regulated by heat in Csspt mutant plants. Thus, these results demonstrated that CsSPT regulated the high-temperature response by recruiting photosynthesis components, signaling pathway molecules, and transcription factors. Our results provide important insights into the heat response mechanism of CsSPT in cucumber and its potential as a target for breeding heat-resistant crops.

Transcriptomic analysis on cucumber tendril reveals GLRs play important roles in thigmotropism and thigmomorphogenesis

Thigmotropism and thigmomorphogenesis are two related and pervasive processes that play crucial roles in plant adaptation to the environment.However,there have been few investigations into the molecular regulatory mechanisms of these phenomena.Cucumber(Cucumis sativus L.)tendrils are ideal material for studying thigmotropism and thigmomorphogenesis because they display a combination of the two processes.Here,we generated the transcriptome profiles of cucumber tendrils at the young,stretch,and coiling stages.Genes related to receptor proteins,transmembrane transport,and ion transport were significantly enriched among those differentially expressed between stages.Pharmacological assays illustrated that three GLUTAMATE RECEPTOR(GLR)genes might play a vital function in perceiving or transducing touch stimulation signals.Comparing the transcriptomes of tendrils and roots after touch stimulation,we found that genes related to extracellular stimulus and xyloglucan metabolism might have conserved functions in the regulation of thigmomorphogenesis.The transcriptome atlas of thigmotropism and thigmomorphogenesis of cucumber tendrils constructed in this study will help further elucidate the molecular mechanisms behind these processes.

The intrinsic developmental age signal defines an age-dependent climbing behavior in cucumber

The tendril is a climbing organ in cucurbits and functions in physical support and to avoid shading by neighboring vegetation.However,how cucurbits produce tendrils to obtain climbing ability is largely unknown.In this study,tendril phenotypes were investigated during different developmental stages.Our results revealed that tendril growth exhibited an age-dependent pattern in cucurbits.Tendril growth was inhibited,and the tendril was formed as a short tendril[nonfunctional tendril(nonF-tendril),approximately 0.1 cm]during the seedling stage.In contrast,enhanced cell proliferation and cell expansion led to rapid elongation of the tendril during the climbing stage,and the tendril formed as a functional tendril(F-tendril,approximately 30 cm)to obtain climbing ability.RT-qPCR detection showed that age-dependent tendril growth correlated negatively with the abundance of the conserved age regulator CsmiR156.Defoliation induced CsmiR156 to inhibit CsSPLs,and F-tendril formation and climbing ability were delayed in defoliated cucumbers,which confirmed the role of CsmiR156 in regulating tendril growth in vivo.Additionally,exogenous gibberellin(GA)treatment showed that GA positively regulated tendril growth,and RT-qPCR detection showed that the GA bio-synthetic genes and metabolic genes were affected by age pathway,suggesting that the age pathway depended on GA bio-synthetic and metabolic pathway to regulate cell expansion to determine tendril growth.In summary,our work reveals that change in tendril type is an important marker of phase transition in cucumber,and tendril growth is regulated by an intrinsic developmental age signal,ensuring that the cucumber obtains climbing ability at a suitable age.

The bHLH transcription factor CsPIF4 positively regulates high temperature-induced hypocotyl elongation in cucumber

High temperature-induced hypocotyl elongation is a typical thermomorphogenesis trait that may significantly affect early seedling growth and subsequent crop yield.The ambient temperature and endogenous auxin are two critical factors that regulate hypocotyl growth.However,the mechanism of temperature and auxin integration in horticultural plants remains poorly understood.In this study,the roles of the basic helix-loop-helix transcription factor CsPIF4 in regulating auxin biosynthesis genes and the auxin content in the hypocotyl of cucumber(Cucumis sativus L.)seedlings under high temperature were investigated.qRT-PCR and in situ hybridization analysis revealed that expression of CsPIF4 was enhanced in the epidermis and vascular bundles in the hypocotyl of cucumber seedlings in response to high temperature.qRT-PCR and HPLC analysis showed that CsPIF4 positively regulated transcription of the auxin biosynthesis gene CsYUC8 and the auxin content in the hypocotyl under high temperature(35℃).The CRISPR/Cas9-mediated knockout of CsPIF4 resulted in a shorter hypocotyl compared with that of the wild type,together with decreased expression of CsYUC8 and lower auxin content in response to high temperature.Furthermore,biochemical assays showed that CsPIF4 could bind directly to the G-box motif of the CsYUC8 promoter and thereby activate CsYUC8 expression.These findings provide insight into the molecular mechanism of high temperature-mediated hypocotyl elongation in cucumber.

A mutation in the promoter of the yellow stripe-like transporter gene in cucumber results in a yellow cotyledon phenotype

Leaf color mutants in higher plants are considered to be ideal materials for studying the chlorophyll biosynthesis,photosynthesis mechanism and chloroplast development.Herein,we identified a spontaneous mutant,yc412,in cultivated cucumber that exhibited yellow cotyledons.The yellow-lethal mutant was diagnosed with an abnormal chloroplast ultrastructure,and reduced photosynthetic capacity and pigment content.Through bulked segregant analysis-based whole-genome sequencing and fine genetic mapping,we narrowed the yellow cotyledons (yc) locus to a 96.8 kb interval on chromosome 3.By resequencing and molecular cloning,we showed that Csyc is a potential candidate gene,which encodes a yellow stripe-like (YSL) transporter.The T to C mutation in the promoter region of Csyc caused the yellow cotyledon phenotype in yc412.Compared to YZU027A (WT),the expression of Csyc was significantly downregulated in the cotyledons of yc412.Silencing of Csyc in cucumber via virus-induced gene silencing resulted in chlorotic leaves,mainly by suppressing the chlorophyll content.Furthermore,a comparative transcriptome analysis revealed that chloroplast-related genes and chlorophyll biosynthesis genes were significantly downregulated in yc412 cotyledons.Our results provide new insights into the molecular function of the YSL transporter in plant chloroplast development and chlorophyll synthesis.

Sea Cucumber Fishery in Aurora, Philippines

This study was conducted to provide a preliminary assessment of the sea cucumber fishery in Aurora, Philippines. This was conducted from February to June 2013. All sea cucumber gatherers and traders, obtained through snowball sampling, in each identified sea cucumber fishery area in six coastal municipalities (Baler, Dipaculao, Dinalungan, Casiguran, Dilasag and Dingalan) were interviewed using a structured interview schedule and focused group discussions (FGD). Results revealed that gathering sea cucumbers were accomplished by handpicking, skin diving or compressor diving. A total of 15 commercial species were regularly gathered. Out of this, three species are of very high economic value (≥Php 4000/kg) (Holothuria nobilis, Holothuria scabra, and Actinopyga lecanora). The average number of kilograms harvested per hour per trip ranges from 0.21 (Holothuria fuscopunctata) to 2.71 (Holothuria scabra), while the average size of commonly gathered sea cucumber varies among species with Holothuria lecanora showed an average length (29.26 cm) greater than recorded 24 cm. Processing mainly involves washing, slicing, boiling, drying and packing. Processed sea cucumbers or trepan were sold to middlemen or direct buyers who usually offered higher prices (36.36% up to 69.20%) than the former. Problems encountered by sea cucumber gatherers and traders include a decline in total catch, a shift in area of collection from intertidal zones to deeper waters, an increase in the number of hours spent in finding and gathering sea cucumbers, the use of compressors in gathering, the decline of larger species and gathering of smaller sizes in huge amount, the absence of conservation program for sea cucumber, and the lenient implementation of government policies with regards to coastal resources management.

Construction of Fusion Expression Vector of α-galactosidase-EGFP in Cucumber

[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter sequence and the coding region of EGFP were amplified by polymerase chain reactions （PCR） with vector pCambia 1303 as the template.Using reverse transcript PCR technology,with total RNAs of cucumber as template,the coding region of acid α-galactosidase Ⅰ in cucumber was amplified.The above three fragments were inserted into the multiple cloning sites of expression vector pCambia 1381c.The fusion expression vector of α-galactosidase-EGFP located at the C-terminal of the target genes was constructed.[Result] After enzyme digestion and sequencing,the fusion expression of α-galactosidase-EGFP in cucumber was constructed successfully.[Conclusion] The research laid the experimental basis for further study on the subcellular localization of α-galactosidase in cucumber.

Effects of Different Irrigation Amounts on Water Consumption and Water Use Efficiency of Greenhouse Cucumber

[Objective] This study was to investigate the effects of different irrigation amount on water consumption and water use efficiency of greenhouse cucumber.[Method]Under the condition of drip irrigation with different water amounts in sunlight greenhouse of the arid areas in Ningxia,the soil water was measured and the water consumption of crop was calculated.[Result]When irrigation amount was 563 mm,the water consumption as a whole gradually increased with the delay of growth period,reached peak during the thriving stage of fruit setting,and then gradually declined;in each treatment,the daily water consumption increased with the increasing of irrigation amount during each growth period.However,the consumption of soil moisture reduced with the significant increase of irrigation.563 mm of irrigation amount could meet the water requirements of cucumber and began to add water to soil,and water utilization efficiency could reach 33.4 kg/m3.[Conclusion]The research had provided theoretical basis for water management in the production process of greenhouse cucumber.

Virus Movement Protein Gene Mediated Resistance Against Cucumber Mosaic Virus Infection

Tobacco ( Nicotiana tabacum L.) “NC89” plants were transformed with deletion mutant of cucumber mosaic virus (CMV) movement protein (MP) gene and full_length CMV MP gene, respectively. The transformed plants were analyzed with polymerase chain reaction (PCR), PCR_Southern, Southern and Western blots. R 0 generation of the transgenic plants were inoculated with CMV. Five out of 10 lines of tobacco plants (BMPK) transformed with CMV MP deletion mutant gene showed high resistance to CMV infection and remained symptomless for up to 50 days post_inoculation. In contrast, tobacco plants (BMPR) transformed with full_length CMV MP gene did not show resistance to CMV infection. However, most of the infected full_length CMV MP gene transgenic plants recovered by showing none or very mild mosaic symptoms in 40 days post_inoculation. The results of R 1 generation of the BMPK transgenic plants tested under field conditions showed that all 5 lines of transgenic plants could delay the virus disease development.

ACC Synthase Gene (ACSG) as a Possible Molecular Marker for Female Lines in Cucumber

利用本实验室根据已知序列分离得到的ACC合酶基因 (ACSG)为探针对不同黄瓜 (CucumissativusL .)品种(系 )的基因组DNA进行Southern杂交 ,初步分析了该基因与黄瓜性别表型之间的相关性。发现在所检测的 10个不同品系中 ,ACSG与雌性系表型之间存在明显的相关关系 ,而且这种相关关系在不同的实验中具有良好的重复性。ACSG基因可能是鉴定黄瓜雌性系的一个分子标记。

Evaluation of Seed Oil Content in Cucumber Germplasm

With 35 different ecological types of cucumber germplasm resources as test materials, the soxhlet extraction was used to extract the crude fat from the cu- cumber seeds to analyze the seed oil content, with the aim to lay foundations for the application and new variety breeding related with cucumber seed oil. The results showed that there were significant differences in seed oil content between different cucumber varieties with the amplitude of variation rangeing from 31.03% to 40.59%, among which the seed oil content of south China cucumber was the highest of 40.59%, while the seed oil content of north China cucumber was the lowest of 31.03%. Four va.rieties with oil content of greater than 38.50% and four varieties with oil content of less than 32.01% werescreened out.

Study on the Activity of Herbal Extract to Kill Root-knot Nematode of Cucumber Seedlings Stage

Present mature plants hydroponic technology was used,combined with some excellent characteristics,such as growth conditions was easy to control and process of root growth was easy to continuously observe,the nematicidal activity of 5 kinds of Chinese herbs extracts and the compound solution of Avermectin,with strong contact toxicity effect indoor,was systematically studied and investigated the affection on the root-knot nematode parasitized on the cucumber seeding stage. It is found that under the premise of no influence on root growth of cucumber,extracts from Picrorhiza scrophulariiflora and Punica granatum showed strong prevention and nematicidal activity,and had the similar efficacy of Avermectin; while the extracts from Cibotium barometz,Aucklandia lappa Decne and Fructus cnidii showed low nematicidal activity and various degrees inhibition effect on plant growth.

Construction of Cucumber Powdery Mildew Early Warning System in Solar Greenhouse Based on Internet of Things

ln order to explore the design and construction of cucumber powdery mildew warning system in solar greenhouse, internet of things technology was used to conduct the real-time dynamic monitoring of the incidence of cucumber powdery mildew and cucumber growth environment in solar greenhouse. The growth environ-ment included temperature and humidity of air and soil. Logistic regression model was used to construct cucumber powdery mildew warning model. The results showed that humidity characteristic variable （maximum air humidity） and temperature characteristic variable （maximum air temperature） had significant effects on the inci-dence probability of cucumber powdery mildew in solar greenhouse. And it was fea-sible to construct cucumber powdery mildew warning system in solar greenhouse with internet of things.

Resistance -induced Effect of Several Chemical Reagents on Cucumber against Pseudoperonospora cubensis

[ Objectlve] The paper was to compare the induced resistance of several chemicals on cucumber against Pseudoperonospora cubensis [ Method ] When the cucumber seedlings grew to 4-leaf stage, the second true leaf was treated with salicylic acid, oxalate, chitosan, calcium nitrate and Na2HPO4, the activity of protective enzymes in the third leaf was determined after 7 d. At the same time, inoculation test was also conducted after induced treatment, and the disease condi- tion was coumed. [ Result ] These chemical treatments promoted protective enzyme activities （POD, SOD, PAL, PPO） in different degree, improved the ultra-weak luminescence, and reduced the disease index of cucumber leaves. [ Conclusion] Five chemical treatments expressed good effects on induced resistance. Among the treatments, oxalate had the best induced effect, followed by salicylic acid, chitosan, Na2 HPO4 and calcium nitrate.

Control Effect of Vegetable Oil on Cucumber Powdery Mildew

[Objective] The study discussed the indoor activity and field control effect of vegetable oil on cucumber powdery mildew.[Method] The cucumber seedlings of Changchunmici were adopted as the material,which were inoculated with the fungus pathogen of cucumber powdery mildew naturally occurred in the field.The indoor protective test and field test were performed respectively to observe the control effects of different plant oils EC on the cucumber powdery mildew.[Result] When six plant oils EC including cottonseed oil,soybean oil,canola oil,corn oil,sesame oil and sunflower oil was diluted into 10 and 5 ml/L,they had better control effect on cucumber powdery mildew.As for the indoor protective effect,the protective effect of the vegetable oil EC was equivalent to the control agent triadimefon EC,the field control effect of vegetable oil EC was in the range of 60%-75%,which was consistent with 69%-70% control effect of contrast agent with 1 ml/L triadimefon EC dilution.[Conclusion] All vegetable oils EC had significant control effect on cucumber powdery mildew in the test.

Effects of Exogenous Glycine Betaine on Oxidation Metabolism in Cucumbers during Low-temperature Storage

[Objective] This study aimed to analyze the effects of different concentrations of glycine betaine（GB） on oxidation metabolism in cucumbers under low-temperature stress and to investigate the possible mechanism of low-temperature resistance in cucumber during low-temperature storage. [Method] Cucumber cultivar Zhongnong No.8 was treated with 0, 5, 10 and 15 mmol/L GB solutions for 15 min and stored at 4 ℃. Changes in oxidative metabolism-related parameters were observed. [Result] Increasing exogenous GB concentration could enhance GB content in cucumbers, decline lipoxygenase（LOX） activity, improve peroxidase（POD） and catalase（CAT） activities, remove effectively hydrogen peroxide（H2O2） and reduce the accumulation of malondialdehyde（MDA）. [Conclusion] Treating cucumbers with10 mmol/L GB exhibited the most remarkable effect.

Breeding of a New Cucumber F_1 Hybrid ‘Oriental Show’

‘Oriental Show’ is a generation of new cucumber variety, with the superi-or inbred line ‘101’ as the female parent and inbred line ‘102’ as the male par-ent. The growth vigor tends to be strong and the first female blossom produces on the 4th or the 5th internode. Fruit is 34 cm long and 32 cm in transverse diameter. Per fruit weighs about 230 g. lt is dark green, densely covered with thorns. lt tastes crisp, containing 167.7 mg/kg Vc, and it is resistant to downy mildew, pow-dery mildew and bacterial angular leaf spot disease. lt is yielding and tolerant to low temperature and weak luminance. The yield can be as high as over 112 443.778 kg/hm^2, and ‘Oriental Show’ is suitable to be cultivated in greenhouses in early spring or solar greenhouses in north China or Henan province.

Effects of Reducing Mn^(2+),Cu^(2+) and Zn^(2+) Concentrations in Nutrient Solution on SOD Activity in Grafted Cucumber Seedling Leaves Under Cold Stress

[Objective] The research aimed to study the effects of reducing the concentrations of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution on the contents of Mn, Cu and Zn of grafted cucumber leaves as well as on the SOD and isozymes activities and the rate of electrolytic leakage under cold stress. [Method] Nutrient solution containing low concentrations of Mn2＋, Cu2＋ or Zn2＋ were used for cultivating the grafted cucumber seedlings for the cold treatment. [Result] The results indicated that reducing the concentrations of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution had declined the SOD activities in grafted cucumber leaves and increased the rates of electrolytic leakage. Mn2＋ had shown the maximum influences, followed by Cu2＋, and Zn2＋ had shown the minimum influences. [Conclusion] Cold resistance of cucumber can be improved by changing the contents of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution.

Effects of Waterlogging Stress on Growth and Physiological Index in Cucumber Seedlings

The effects of waterlogging stress on the growth, chlorophyll and MDA content of cucumber seedlings were studied by using cucumber varieties （Cucumis sativus L. cv. Jinyan No.7） as materials. The results show that plant height, leaf number, shoot fresh weight, total root length, total projected area and total surface area of cucumber seedlings showed insignificant differences with those of the control; root fresh weight, root average diameter, total root volume were significantly higher, but the number of root tips and furcation number increased first and then reduced; the chlorophyll content of cucumber seedlings was significantly lower than that of control; MDA content was significantly higher than that of control under waterlogging stress. It was showed that the cucumber was tolerant to waterlogging stress, but the ability of waterlogging stress was limited, and the physiological was significantly hurt and photosynthesis of cucumber seedlings was significantly affected by waterlogging stress.

Effects of Water Stress on the Mssbauer Spectrum of Fe^(2+) /Fe^(3+) Components in Cucumber PSⅡ Particles

There were four doublets in the Mssbauer spectrum of cucumber (Cucumis sativus L.) photosystemⅡ (PSⅡ) particles. According to the value of isomer shift and quadrupole splitting, they represented oxidized cytochrome b559 (Cyt_b559), reduced Cyt_b559, Fe 3+ _quinone (Q) complex, and Fe 2+ _Q complex respectively. After water_stress, the electron transport rate between Q A (primary quinone electron acceptor of PSⅡ)/Q B(secondary quinone electron acceptor of PSⅡ) was affected and the absorption doublets of Fe 2+ disappeared, suggesting that the reduced Cyt_b559 and Fe 2+ _Q complex had been oxidized. The results indicated that water_stress had changed the redox status of iron in the iron_quinone complex. Iron took part in electron transport through the change from a state of reduction to oxidation.