PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular...PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular mass of 37.1 kDa, interrupted by one intron of 95 bp in, length. Sequence analysis revealed that it has five potential N-giycosylation sites, two protein kinase C phosphrylation sites, five casin kinase Ⅱ phosphrylation sites and four N-myristoylation sites. The amino acids sequences alignment confirmed that ^145 LRR stucture was highly conserved in all aligned PGIP sequences.展开更多
The purpose of the present studies was analysis of the age induced changes in photochemical efficiency and xanthophyils cycle pigments of the primary cabbage (Brassica oleracea L. cv. Capitata f. alba) leaves. Photo...The purpose of the present studies was analysis of the age induced changes in photochemical efficiency and xanthophyils cycle pigments of the primary cabbage (Brassica oleracea L. cv. Capitata f. alba) leaves. Photochemical efficiency of photosystem Ⅱ (PS Ⅱ) was studied by a pulse amplitude modulated chlorophyll fluorescence apparatus, chlorophyll concentration was analysis spectrophotometrically and xanthophyll cycle pigments were estimated by high-pressure liquid chromatography (HPLC). Leaf senescence was accompanied with a decrease both in chlorophylls concentration, the photochemical efficiency and rate constant for PS Ⅱ photochemistry whereas non-photochemical parameters increased. Excitation pressure (1-qP) which is a measure of relative lumen acidification increased by 1.2x in aging leaves. The maximum quantum yield of PS Ⅱ showed no significant change. The level of de-epoxidised xanthophylls increased but the concentration of mono- and di-epoxy xanthophylls decreased in aging leaves. A linear relationship between the excitation pressure and the depoxidation state of the xanthophyll cycle pigments and lutein, during the onset of senescence suggests that excitation pressure can be used as a sensor for monitoring the onset of senescence as well for the de-epoxidation state of the xanthophylls responsible for non-photochemical quenching in stressed leaves.展开更多
Cowpea Trypsin Inhibitor (CpTI) gene was transferred into the cotyle dons and hypocotyls of cauliflower by Agrobacterium-mediated transformation met hod. The best selective concentration of kanamycin (kan) was 15 mg L...Cowpea Trypsin Inhibitor (CpTI) gene was transferred into the cotyle dons and hypocotyls of cauliflower by Agrobacterium-mediated transformation met hod. The best selective concentration of kanamycin (kan) was 15 mg L-1. The con centration of carbencillin (carb) was 500 mg L-1. 14 transgenic cauliflower pla nts were obtained. The putative transformants were assayed by PCR and Southern b lotting analysis. The results indicated that CpTI gene was transferred into caul iflower successfully.展开更多
SSR analysis on genetic diversity of 30 samples was carried out. Five primers selected from 36 primers were used to amplify 30 samples in this experiment, PCR products were separated by 6% polyacrylamide gel electroph...SSR analysis on genetic diversity of 30 samples was carried out. Five primers selected from 36 primers were used to amplify 30 samples in this experiment, PCR products were separated by 6% polyacrylamide gel electrophoresis, silver staining and photographed. The results of SSR were analyzed by UPGMA clustering. The results showed that a total of 21 gene alleles were detected by 5 SSR primers. The number of alleles ranged from 2 to 5 with an average of 4.2.PIC range was 0.257-0.92 1, with an average of 0.543. The average coefficient of genetic similarity of SSR markers among materials was 0.432. Some of cabbage cultivars in the experiment were divided into four groups except cultivars which come from Japan.展开更多
We conducted plant growth experiments in microbe-free vermiculite to study the effects of four types of fermented seaweed liquid fertilizer (SLF) made from nori (Pyropia yezoensis) seaweed on the germination, plant gr...We conducted plant growth experiments in microbe-free vermiculite to study the effects of four types of fermented seaweed liquid fertilizer (SLF) made from nori (Pyropia yezoensis) seaweed on the germination, plant growth characteristics, SPAD value, and nutrient content and uptake of komatsuna (Brassica rapa L. var. wakana komatsuna). The four types of fermented nori SLF used in this study were prepared by anaerobic fermentation of unwashed nori (SLF1), aerobic fermentation of unwashed nori (SLF2), anaerobic fermentation of washed nori (SLF3), and aerobic fermentation of washed nori (SLF4). Komatsuna seeds treated with 200-, 300-, and 400-fold dilutions of SLFs exhibited improved relative germination ratios (RGRs) at 3 and 4 days after sowing (DAS). At 4 DAS, the RGRs of seeds treated with 10-, 100-, 200-, 300-, and 400-fold SLF dilutions showed no differential effect. Seeds treated with undiluted SLFs did not germinate by 4 DAS. SLF1 may promote komatsuna seed germination. The nitrogen (N), calcium, magnesium, sodium (Na), and iodine (I) contents of plants treated with SLF1 were significantly increased relative to plants treated with the other SLFs. Moreover, the I and Na contents of plants were significantly increased by foliar spray application of different dilutions of SLF1. However, SLF treatment markedly reduced the shoot dry weight compared with 1/2-strength modified Hoagland nutrient (MHN) solution, although the same amounts of N and K were applied. SPAD values of the plants treated with SLFs were significantly higher than those of plants treated with MHN. Foliar treatment with SLFs had no significant effect on plant growth, SPAD value, or uptake of nutrients (except Na) relative to the control, but the I content was increased. Plants treated with SLF1 and SLF2 exhibited the highest Na uptake. Foliar spray treatments with SLF1 resulted in the highest I contents in plants. Based on our results, SLF1 is suitable for use as a liquid fertilizer to promote germination and increase nutrient content in komatsuna. These results need to be followed up in soil experiments in the presence of microbes in the rhizosphere.展开更多
Many of the extraction and amino acid analysis methodologies currently employed do not provide complete analysis of all the physiological amino acids and biogenic amines. Extraction procedures frequently employ dilute...Many of the extraction and amino acid analysis methodologies currently employed do not provide complete analysis of all the physiological amino acids and biogenic amines. Extraction procedures frequently employ dilute acid which partially converts gln and asn to glu and asp. A commonly used pre-column derivatizing agent, o-phthalaldehyde, does not react with the imino acids, pro and hydroxypro. The purpose of this investigation was to integrate extraction and analysis procedures into a reliable method for measuring the complete physiological amino acid profiles of fruit and vegetables using HPLC instrumentation commonly available to most laboratories. Water extraction of ground, frozen-thawed tissues effected complete recovery of the physiological amino acids as demonstrated by spiking experiments and tissue combination experiments. HPLC of dabsyl derivatives of the free amino acids allowed their quantification in a selection of fruit and vegetables. Physiological amino acid levels were determined for peach, apple, potato, onion, tomato, bell pepper, broccoli, and seven types of cucurbits. The coefficient of variation for estimation of an amino acid level generally fell in the range of 5% to 7%. Because of marked variability in physiological amino acid content as a result of growing conditions, cultural practices, and inherent cultivar differences, comparisons of results with literature values were not possible.展开更多
Cytokinins are a class of phytohormones that promote cell division and differentiation and are thought to affect plant immunity to multiple pathogens.However,a comprehensive analysis of cytokinin dehydrogenase/oxidase...Cytokinins are a class of phytohormones that promote cell division and differentiation and are thought to affect plant immunity to multiple pathogens.However,a comprehensive analysis of cytokinin dehydrogenase/oxidase(CKX)family genes in cabbage has not been reported.In this study,a total of 36 CKX genes were identified using a genome-wide search method.Phylogenetic analysis classified these genes into three groups.They were distributed unevenly across nine chromosomes in B.oleracea,and 15 of them did not contain any introns.The results of colinearity analysis showed that 36 CKX gene in Arabidopsis was present in several copies in the Brassica oleracea genome.An analysis of cisacting elements indicated that all genes possessed at least one stress or hormone responsive cis-acting element.A heatmap of CKX gene expression showed the patterns of expression of these genes in various tissues and organs.Three genes(Bol028363,Bol031036 and Bol018140)were relatively highly expressed in all of the investigated tissues under normal conditions,showing the expression profile of housekeeping genes.Generally,the expression patterns of CKX genes in Jingfeng 1 and Xiangan 336 were quite different under the same treatment.Notably,three genes(Bol020547,Bol028392 and Bol045724)were significantly down-regulated and up-regulated in the susceptible and resistant material,respectively,after inoculation,which may indicate their crucial roles in resistance to clubroot disease.The results provide insights for better understanding the roles of CKX genes in the B.oleracea–P.brassicae interaction.展开更多
Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plan...Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker(50 rpm,21℃,3h,2500 lux light),and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light(4000 lux)for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere展开更多
Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growt...Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growth rate,dry weight,fresh weight and relative water content of the plants were determined. After 6h of treatment,the activity and gene expression of three antioxidant enzymes,superoxide dismutase( SOD),catalase( CAT) and ascorbate peroxidase( APX) in plants,were measured. The results showed that the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in collard seedlings were higher in the treatment of salt stress with the addition of 0. 05 mmol/L exogenous H_2O_2 than in the simple salt stress treatment; and when endogenous H_2O_2 was removed,the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in plant seedlings were lower than those under simple salt stress. It is speculated that under salt stress,H_2O_2 is involved in the regulation of antioxidant defense gene expression,and it might be an important regulator of salt-induced antioxidant system in collard leaves.展开更多
To better understand waterlogging effect on leaf senescence in winter rapseed (Brassica napus L.) during flowering stage, experiments were designed to explore foliar K application influences on adverse effects of wate...To better understand waterlogging effect on leaf senescence in winter rapseed (Brassica napus L.) during flowering stage, experiments were designed to explore foliar K application influences on adverse effects of waterlogging stress. Winter rapeseed was sprayed with K after waterlogging at initial flowering stage. Results indicated that waterlogging significantly decreased leaf net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci) and transpiration rate (Tr). It also declined maximum quantum yield of PS II (Fv/Fm), quantum yield of electron transport (ΦPS II) and photochemical quenching (qP), but increased leaf non-photochemical quenching (NPQ) and minimal fluorescence (Fo). Interestingly, exogenous application of K significantly alleviated waterlogging-induced photosynthesis inhibition. Foliar K application increased RuBisCO activation, chlorophyll and soluble protein contents, while significantly decreased MDA content under waterlogging stress. Moreover, K supplementation improved accumulation of K+, Ca2+, Mg2+, N, Zn2+, Mn2+, Fe2+ in leaves. In general, foliar K application is effective inalleviating deleterious effects of waterlogging stress and delays leaf senescence of winter rapeseed.展开更多
We have done a comparative study of ion status, growth and biochemical parameters in shoots and roots of seablite (Suaeda altissima (L.) Pall.) and spinach (Spinacia oleracea L.) grown with different salinity levels i...We have done a comparative study of ion status, growth and biochemical parameters in shoots and roots of seablite (Suaeda altissima (L.) Pall.) and spinach (Spinacia oleracea L.) grown with different salinity levels in the medium (0.5 - 750 mМ). A distinctive feature of the halophyte was a high Na+ content in tissues at its low concentration in the medium (0.5 mM). In these conditions, Na+ accumulation in seablite roots was four-fold higher than in spinach roots, and Na+ content in seablite leaves was almost 20-fold higher than in spinach. Together with an increase in sodium concentration in the medium, K+ content decreased six-fold in seablite leaves, while in spinach it did not decrease so drastically. We can suppose that in the halophyte, some processes occur only in the presence of sodium, and these functions of sodium cannot be fully fulfilled by potassium. Analysis of protein and total nitrogen content in tissues shows that at high salinity, the ability to synthesize non-protein nitrogen-containing compounds increases in the halophyte and decreases in the glycophyte. Data on proline content dynamics show that its increase in tissues of spinach (salinity levels 150 and 250 mМ) and seablite (salinity levels 0.5 and 750 mМ) is an indicator of plant injury. In seablite and spinach, proline is not a major osmoregulator. Its concentration both in roots and leaves was no more than 2.5 μmol/g fresh weight. The data presented in this work concern the accumulation and distribution of Na+, Cl?, K+ and ions, as well as growth and biochemical parameters. Our data show that the development of adaptation reactions in the whole plants in the conditions of high salinity is determined by morphofunctional systems and their interaction.展开更多
Brassica oleracea has been developed into many important crops,including cabbage,kale,cauliflower,broccoli and so on.The genome and gene annotation of cabbage(cultivar JZS),a representative morphotype of B.oleracea,ha...Brassica oleracea has been developed into many important crops,including cabbage,kale,cauliflower,broccoli and so on.The genome and gene annotation of cabbage(cultivar JZS),a representative morphotype of B.oleracea,has been widely used as a common reference in biological research.Although its genome assembly has been updated twice,the current gene annotation still lacks information on untranslated regions(UTRs)and alternative splicing(AS).Here,we constructed a high-quality gene annotation(JZSv3)using a full-length transcriptome acquired by nanopore sequencing,yielding a total of 59452 genes and 75684 transcripts.Additionally,we re-analyzed the previously reported transcriptome data related to the development of different tissues and cold response using JZSv3 as a reference,and found that 3843 out of 11908 differentially expressed genes(DEGs)underwent AS during the development of different tissues and 309 out of 903 cold-related genes underwent AS in response to cold stress.Meanwhile,we also identified many AS genes,including BolLHCB5 and BolHSP70,that displayed distinct expression patterns within variant transcripts of the same gene,highlighting the importance of JZSv3 as a pivotal reference for AS analysis.Overall,JZSv3 provides a valuable resource for exploring gene function,especially for obtaining a deeper understanding of AS regulation mechanisms.展开更多
Low temperature is one of the most important abiotic factors inhibiting growth, productivity, and distribution of rapeseed(Brassica napus L.). Therefore, it is important to identify and cultivate cold-tolerant germpla...Low temperature is one of the most important abiotic factors inhibiting growth, productivity, and distribution of rapeseed(Brassica napus L.). Therefore, it is important to identify and cultivate cold-tolerant germplasm. The objective of this study was to figure out the mechanism of chilling(4 and 2°C) and freezing(–2 and –4°C) stresses along with a control(22°C) in B. napus cultivars(1801 and C20) under controlled environment(growth chamber). The experiment was arranged in a complete randomized design with three replications. Our results exhibited that under chilling and freezing stresses, the increment of proline accumulation, soluble sugar and protein contents, and antioxidant enzyme activity were enhanced more in 1801 cultivar compared with C20 cultivar. At –2 and –4°C, the seedlings of C20 cultivar died completely compared with 1801 cultivar. Hydrogen peroxide(H2 O2) and malondialdehyde contents(MDA) increased in both cultivars, but when the temperature was decreased up to –2 and –4°C, the MDA and H2 O2 contents continuously dropped in 1801 cultivar. Moreover, we found that leaf abscisic acid(ABA) was enhanced in 1801 cultivar under chilling and freezing stresses. Additionally, the transcriptional regulations of cold-tolerant genes(COLD1, CBF4, COR6.6, COR15, and COR25) were also determined using real-time quantitative PCR(RT-q PCR). RT-q PCR showed that higher expression of these genes were found in 1801 as compared to C20 under cold stress(chilling and freezing stresses). Therefore, it is concluded from this experiment that 1801 cultivar has a higher ability to respond to cold stress(chilling and freezing stresses) by maintaining hormonal, antioxidative, and osmotic activity along with gene transcription process than C20. The result of this study will provide a solid foundation for understanding physiological and molecular mechanisms of cold stress signaling in rapeseed(B. napus).展开更多
To better understand the effect of high temperature on seed viability, artificial aging treatments were implemented on 472 accessions of rapeseed (Brassica napus L.) by constant high temperature at 70℃for 8h. Results...To better understand the effect of high temperature on seed viability, artificial aging treatments were implemented on 472 accessions of rapeseed (Brassica napus L.) by constant high temperature at 70℃for 8h. Results showed a remarkable impact of constant heat on seed germination. After heat treatment, considerable variation was found in seed germinability, some genotypes even lost the ability to germinate. The effect of accelerated aging was highly significant. Germination parameters varied among ecotypes. Seed viability varied with different origin, and was significantly and positively correlated with seed yield per plant and dry weight of aboveground parts per plant among 14 agronomic traits. Germination traits were positively correlated with oil contents.展开更多
Effective silique number per plant(ESN), seeds per silique(SS), thousand seeds weight(TSW), silique length(SL) and silique density(SD) are important seed yield potential determinant traits in rapeseed(Brassica napus L...Effective silique number per plant(ESN), seeds per silique(SS), thousand seeds weight(TSW), silique length(SL) and silique density(SD) are important seed yield potential determinant traits in rapeseed(Brassica napus L.), which are controlled by quantitative trait loci(QTL). Mapping QTL to delimited chromosomal region offers an effective method for genetic dissection of these traits. A set of 96 double haploid(DH) lines were developed by crossing 2 Brassica napus lines R1 and R2, and an immortalized F_2(IF_2) population containing 124 combinations was developed by crossing those DH lines. DH populations were planted at 2 locations for 2 years and IF_2 populations were planted in 2 locations for 1 year. Based on the established 2,217.2 cM length high density genetic map, 42 QTLs were identified, with 26 QTLs detected repeatedly in different environments or populations, including 8 for SL, 7 for TSW, 4 for ESN, 4 for SS and 3 for SD. Among these identified QTLs, 3, 4, 1, 1 and 3 QTLs were considered as major QTLs for SL, TSW, ESN, SS and SD, respectively. In addition, 2 QTLs on A9 chromosome which control multiple traits were identified. These results warrant further study of fine mapping for yield and yield components.展开更多
基金Supported by School High-level Talent Starting Fund of Qingdao Agriculture University:Studies on Clone and Evolution of PGIPGene from Brassica crops(630745)~~
文摘PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular mass of 37.1 kDa, interrupted by one intron of 95 bp in, length. Sequence analysis revealed that it has five potential N-giycosylation sites, two protein kinase C phosphrylation sites, five casin kinase Ⅱ phosphrylation sites and four N-myristoylation sites. The amino acids sequences alignment confirmed that ^145 LRR stucture was highly conserved in all aligned PGIP sequences.
文摘The purpose of the present studies was analysis of the age induced changes in photochemical efficiency and xanthophyils cycle pigments of the primary cabbage (Brassica oleracea L. cv. Capitata f. alba) leaves. Photochemical efficiency of photosystem Ⅱ (PS Ⅱ) was studied by a pulse amplitude modulated chlorophyll fluorescence apparatus, chlorophyll concentration was analysis spectrophotometrically and xanthophyll cycle pigments were estimated by high-pressure liquid chromatography (HPLC). Leaf senescence was accompanied with a decrease both in chlorophylls concentration, the photochemical efficiency and rate constant for PS Ⅱ photochemistry whereas non-photochemical parameters increased. Excitation pressure (1-qP) which is a measure of relative lumen acidification increased by 1.2x in aging leaves. The maximum quantum yield of PS Ⅱ showed no significant change. The level of de-epoxidised xanthophylls increased but the concentration of mono- and di-epoxy xanthophylls decreased in aging leaves. A linear relationship between the excitation pressure and the depoxidation state of the xanthophyll cycle pigments and lutein, during the onset of senescence suggests that excitation pressure can be used as a sensor for monitoring the onset of senescence as well for the de-epoxidation state of the xanthophylls responsible for non-photochemical quenching in stressed leaves.
文摘Cowpea Trypsin Inhibitor (CpTI) gene was transferred into the cotyle dons and hypocotyls of cauliflower by Agrobacterium-mediated transformation met hod. The best selective concentration of kanamycin (kan) was 15 mg L-1. The con centration of carbencillin (carb) was 500 mg L-1. 14 transgenic cauliflower pla nts were obtained. The putative transformants were assayed by PCR and Southern b lotting analysis. The results indicated that CpTI gene was transferred into caul iflower successfully.
文摘SSR analysis on genetic diversity of 30 samples was carried out. Five primers selected from 36 primers were used to amplify 30 samples in this experiment, PCR products were separated by 6% polyacrylamide gel electrophoresis, silver staining and photographed. The results of SSR were analyzed by UPGMA clustering. The results showed that a total of 21 gene alleles were detected by 5 SSR primers. The number of alleles ranged from 2 to 5 with an average of 4.2.PIC range was 0.257-0.92 1, with an average of 0.543. The average coefficient of genetic similarity of SSR markers among materials was 0.432. Some of cabbage cultivars in the experiment were divided into four groups except cultivars which come from Japan.
文摘We conducted plant growth experiments in microbe-free vermiculite to study the effects of four types of fermented seaweed liquid fertilizer (SLF) made from nori (Pyropia yezoensis) seaweed on the germination, plant growth characteristics, SPAD value, and nutrient content and uptake of komatsuna (Brassica rapa L. var. wakana komatsuna). The four types of fermented nori SLF used in this study were prepared by anaerobic fermentation of unwashed nori (SLF1), aerobic fermentation of unwashed nori (SLF2), anaerobic fermentation of washed nori (SLF3), and aerobic fermentation of washed nori (SLF4). Komatsuna seeds treated with 200-, 300-, and 400-fold dilutions of SLFs exhibited improved relative germination ratios (RGRs) at 3 and 4 days after sowing (DAS). At 4 DAS, the RGRs of seeds treated with 10-, 100-, 200-, 300-, and 400-fold SLF dilutions showed no differential effect. Seeds treated with undiluted SLFs did not germinate by 4 DAS. SLF1 may promote komatsuna seed germination. The nitrogen (N), calcium, magnesium, sodium (Na), and iodine (I) contents of plants treated with SLF1 were significantly increased relative to plants treated with the other SLFs. Moreover, the I and Na contents of plants were significantly increased by foliar spray application of different dilutions of SLF1. However, SLF treatment markedly reduced the shoot dry weight compared with 1/2-strength modified Hoagland nutrient (MHN) solution, although the same amounts of N and K were applied. SPAD values of the plants treated with SLFs were significantly higher than those of plants treated with MHN. Foliar treatment with SLFs had no significant effect on plant growth, SPAD value, or uptake of nutrients (except Na) relative to the control, but the I content was increased. Plants treated with SLF1 and SLF2 exhibited the highest Na uptake. Foliar spray treatments with SLF1 resulted in the highest I contents in plants. Based on our results, SLF1 is suitable for use as a liquid fertilizer to promote germination and increase nutrient content in komatsuna. These results need to be followed up in soil experiments in the presence of microbes in the rhizosphere.
文摘Many of the extraction and amino acid analysis methodologies currently employed do not provide complete analysis of all the physiological amino acids and biogenic amines. Extraction procedures frequently employ dilute acid which partially converts gln and asn to glu and asp. A commonly used pre-column derivatizing agent, o-phthalaldehyde, does not react with the imino acids, pro and hydroxypro. The purpose of this investigation was to integrate extraction and analysis procedures into a reliable method for measuring the complete physiological amino acid profiles of fruit and vegetables using HPLC instrumentation commonly available to most laboratories. Water extraction of ground, frozen-thawed tissues effected complete recovery of the physiological amino acids as demonstrated by spiking experiments and tissue combination experiments. HPLC of dabsyl derivatives of the free amino acids allowed their quantification in a selection of fruit and vegetables. Physiological amino acid levels were determined for peach, apple, potato, onion, tomato, bell pepper, broccoli, and seven types of cucurbits. The coefficient of variation for estimation of an amino acid level generally fell in the range of 5% to 7%. Because of marked variability in physiological amino acid content as a result of growing conditions, cultural practices, and inherent cultivar differences, comparisons of results with literature values were not possible.
基金supported by the Youth Science Fund Project(Grant No. 31801876)。
文摘Cytokinins are a class of phytohormones that promote cell division and differentiation and are thought to affect plant immunity to multiple pathogens.However,a comprehensive analysis of cytokinin dehydrogenase/oxidase(CKX)family genes in cabbage has not been reported.In this study,a total of 36 CKX genes were identified using a genome-wide search method.Phylogenetic analysis classified these genes into three groups.They were distributed unevenly across nine chromosomes in B.oleracea,and 15 of them did not contain any introns.The results of colinearity analysis showed that 36 CKX gene in Arabidopsis was present in several copies in the Brassica oleracea genome.An analysis of cisacting elements indicated that all genes possessed at least one stress or hormone responsive cis-acting element.A heatmap of CKX gene expression showed the patterns of expression of these genes in various tissues and organs.Three genes(Bol028363,Bol031036 and Bol018140)were relatively highly expressed in all of the investigated tissues under normal conditions,showing the expression profile of housekeeping genes.Generally,the expression patterns of CKX genes in Jingfeng 1 and Xiangan 336 were quite different under the same treatment.Notably,three genes(Bol020547,Bol028392 and Bol045724)were significantly down-regulated and up-regulated in the susceptible and resistant material,respectively,after inoculation,which may indicate their crucial roles in resistance to clubroot disease.The results provide insights for better understanding the roles of CKX genes in the B.oleracea–P.brassicae interaction.
文摘Eight F<sub>1</sub>-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker(50 rpm,21℃,3h,2500 lux light),and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light(4000 lux)for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere
基金Supported by Science and Technology Development Planning Project of Henan Province(182102110305)
文摘Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growth rate,dry weight,fresh weight and relative water content of the plants were determined. After 6h of treatment,the activity and gene expression of three antioxidant enzymes,superoxide dismutase( SOD),catalase( CAT) and ascorbate peroxidase( APX) in plants,were measured. The results showed that the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in collard seedlings were higher in the treatment of salt stress with the addition of 0. 05 mmol/L exogenous H_2O_2 than in the simple salt stress treatment; and when endogenous H_2O_2 was removed,the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in plant seedlings were lower than those under simple salt stress. It is speculated that under salt stress,H_2O_2 is involved in the regulation of antioxidant defense gene expression,and it might be an important regulator of salt-induced antioxidant system in collard leaves.
文摘To better understand waterlogging effect on leaf senescence in winter rapseed (Brassica napus L.) during flowering stage, experiments were designed to explore foliar K application influences on adverse effects of waterlogging stress. Winter rapeseed was sprayed with K after waterlogging at initial flowering stage. Results indicated that waterlogging significantly decreased leaf net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci) and transpiration rate (Tr). It also declined maximum quantum yield of PS II (Fv/Fm), quantum yield of electron transport (ΦPS II) and photochemical quenching (qP), but increased leaf non-photochemical quenching (NPQ) and minimal fluorescence (Fo). Interestingly, exogenous application of K significantly alleviated waterlogging-induced photosynthesis inhibition. Foliar K application increased RuBisCO activation, chlorophyll and soluble protein contents, while significantly decreased MDA content under waterlogging stress. Moreover, K supplementation improved accumulation of K+, Ca2+, Mg2+, N, Zn2+, Mn2+, Fe2+ in leaves. In general, foliar K application is effective inalleviating deleterious effects of waterlogging stress and delays leaf senescence of winter rapeseed.
文摘We have done a comparative study of ion status, growth and biochemical parameters in shoots and roots of seablite (Suaeda altissima (L.) Pall.) and spinach (Spinacia oleracea L.) grown with different salinity levels in the medium (0.5 - 750 mМ). A distinctive feature of the halophyte was a high Na+ content in tissues at its low concentration in the medium (0.5 mM). In these conditions, Na+ accumulation in seablite roots was four-fold higher than in spinach roots, and Na+ content in seablite leaves was almost 20-fold higher than in spinach. Together with an increase in sodium concentration in the medium, K+ content decreased six-fold in seablite leaves, while in spinach it did not decrease so drastically. We can suppose that in the halophyte, some processes occur only in the presence of sodium, and these functions of sodium cannot be fully fulfilled by potassium. Analysis of protein and total nitrogen content in tissues shows that at high salinity, the ability to synthesize non-protein nitrogen-containing compounds increases in the halophyte and decreases in the glycophyte. Data on proline content dynamics show that its increase in tissues of spinach (salinity levels 150 and 250 mМ) and seablite (salinity levels 0.5 and 750 mМ) is an indicator of plant injury. In seablite and spinach, proline is not a major osmoregulator. Its concentration both in roots and leaves was no more than 2.5 μmol/g fresh weight. The data presented in this work concern the accumulation and distribution of Na+, Cl?, K+ and ions, as well as growth and biochemical parameters. Our data show that the development of adaptation reactions in the whole plants in the conditions of high salinity is determined by morphofunctional systems and their interaction.
基金supported by the National Natural Science Foundation of China (Grant Nos.31972411,31722048,and 31630068)the Central Public-interest Scientific Institution Basal Research Fund (Grant No.Y2022PT23)+1 种基金the Innovation Program of the Chinese Academy of Agricultural Sciences,and the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture and Rural Affairs,P.R.Chinasupported by NIFA,the Department of Agriculture,via UC-Berkeley,USA。
文摘Brassica oleracea has been developed into many important crops,including cabbage,kale,cauliflower,broccoli and so on.The genome and gene annotation of cabbage(cultivar JZS),a representative morphotype of B.oleracea,has been widely used as a common reference in biological research.Although its genome assembly has been updated twice,the current gene annotation still lacks information on untranslated regions(UTRs)and alternative splicing(AS).Here,we constructed a high-quality gene annotation(JZSv3)using a full-length transcriptome acquired by nanopore sequencing,yielding a total of 59452 genes and 75684 transcripts.Additionally,we re-analyzed the previously reported transcriptome data related to the development of different tissues and cold response using JZSv3 as a reference,and found that 3843 out of 11908 differentially expressed genes(DEGs)underwent AS during the development of different tissues and 309 out of 903 cold-related genes underwent AS in response to cold stress.Meanwhile,we also identified many AS genes,including BolLHCB5 and BolHSP70,that displayed distinct expression patterns within variant transcripts of the same gene,highlighting the importance of JZSv3 as a pivotal reference for AS analysis.Overall,JZSv3 provides a valuable resource for exploring gene function,especially for obtaining a deeper understanding of AS regulation mechanisms.
基金supported by the National Key Research and Development Program of China (2017YFD0101700)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciencesthe Hubei Agricultural Science and Technology Innovation Center, China
文摘Low temperature is one of the most important abiotic factors inhibiting growth, productivity, and distribution of rapeseed(Brassica napus L.). Therefore, it is important to identify and cultivate cold-tolerant germplasm. The objective of this study was to figure out the mechanism of chilling(4 and 2°C) and freezing(–2 and –4°C) stresses along with a control(22°C) in B. napus cultivars(1801 and C20) under controlled environment(growth chamber). The experiment was arranged in a complete randomized design with three replications. Our results exhibited that under chilling and freezing stresses, the increment of proline accumulation, soluble sugar and protein contents, and antioxidant enzyme activity were enhanced more in 1801 cultivar compared with C20 cultivar. At –2 and –4°C, the seedlings of C20 cultivar died completely compared with 1801 cultivar. Hydrogen peroxide(H2 O2) and malondialdehyde contents(MDA) increased in both cultivars, but when the temperature was decreased up to –2 and –4°C, the MDA and H2 O2 contents continuously dropped in 1801 cultivar. Moreover, we found that leaf abscisic acid(ABA) was enhanced in 1801 cultivar under chilling and freezing stresses. Additionally, the transcriptional regulations of cold-tolerant genes(COLD1, CBF4, COR6.6, COR15, and COR25) were also determined using real-time quantitative PCR(RT-q PCR). RT-q PCR showed that higher expression of these genes were found in 1801 as compared to C20 under cold stress(chilling and freezing stresses). Therefore, it is concluded from this experiment that 1801 cultivar has a higher ability to respond to cold stress(chilling and freezing stresses) by maintaining hormonal, antioxidative, and osmotic activity along with gene transcription process than C20. The result of this study will provide a solid foundation for understanding physiological and molecular mechanisms of cold stress signaling in rapeseed(B. napus).
基金supported by the Nature Science Foundation of China (31601341)
文摘To better understand the effect of high temperature on seed viability, artificial aging treatments were implemented on 472 accessions of rapeseed (Brassica napus L.) by constant high temperature at 70℃for 8h. Results showed a remarkable impact of constant heat on seed germination. After heat treatment, considerable variation was found in seed germinability, some genotypes even lost the ability to germinate. The effect of accelerated aging was highly significant. Germination parameters varied among ecotypes. Seed viability varied with different origin, and was significantly and positively correlated with seed yield per plant and dry weight of aboveground parts per plant among 14 agronomic traits. Germination traits were positively correlated with oil contents.
基金supported by the National Key Research and Development Program of China(No.2016YFD0101300)the Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences(Group No.118)+1 种基金the Earmarked Fund for China Agriculture Research System(CARS-12)Hubei Agricultural Science and Technology Innovation Center(201620000001048)
文摘Effective silique number per plant(ESN), seeds per silique(SS), thousand seeds weight(TSW), silique length(SL) and silique density(SD) are important seed yield potential determinant traits in rapeseed(Brassica napus L.), which are controlled by quantitative trait loci(QTL). Mapping QTL to delimited chromosomal region offers an effective method for genetic dissection of these traits. A set of 96 double haploid(DH) lines were developed by crossing 2 Brassica napus lines R1 and R2, and an immortalized F_2(IF_2) population containing 124 combinations was developed by crossing those DH lines. DH populations were planted at 2 locations for 2 years and IF_2 populations were planted in 2 locations for 1 year. Based on the established 2,217.2 cM length high density genetic map, 42 QTLs were identified, with 26 QTLs detected repeatedly in different environments or populations, including 8 for SL, 7 for TSW, 4 for ESN, 4 for SS and 3 for SD. Among these identified QTLs, 3, 4, 1, 1 and 3 QTLs were considered as major QTLs for SL, TSW, ESN, SS and SD, respectively. In addition, 2 QTLs on A9 chromosome which control multiple traits were identified. These results warrant further study of fine mapping for yield and yield components.