1.Study on pollen viability and stigma receptivity of 5 Iris plants.
Xiaofang YU ; Haiqin ZHANG ; Tianyi JIANG ; Yonghong ZHOU
China Journal of Chinese Materia Medica 2009;34(10):1204-1206
OBJECTIVETo increase the reproduction efficiency of Iris plants.
METHODPollen viability, stigmatic receptivity, the color of anther and stigma of 5 Iris plants were observed during blooming.
RESULT1. The highest pollen viability was in 4 hours after blooming; 2. The stigmatic receptivities of I. sichuanensis, I. leptophylla, I. lactea and I. goniocarpa were strong in 4 hours after blooming, while that of I. lactea var. chinensis was strong in 2 hours after blooming; 3. The color of anther could reflect the pollen viability, but could not indicate the viability level; 4. The stigma color could not reflect the receptivity of stigma.
CONCLUSIONThe optimum artificial pollination time of these five species were 12:00 -14:00.
Biological Evolution ; Color ; Flowers ; physiology ; Iris Plant ; physiology ; Odorants ; Plant Infertility ; Pollen ; growth & development ; Pollination ; physiology ; Reproduction ; physiology ; Species Specificity
2.Characteristics and abortion cytological pathways of male sterility of Bupleurum chinense.
Chengmin YANG ; Jing YU ; Jianhe WEI ; Chun SUI ; Qinglong CHU
China Journal of Chinese Materia Medica 2010;35(24):3246-3249
OBJECTIVETo study the characteristics of male sterility of Bupleurum chinense and further explore the developmental period and reason of abortion for the male sterile plants.
METHODThe morphological characteristics of B. chinense male sterile and normal plants were investigated and compared. The anther development process and pollen viability of two types of plants were examined by microscopic assay.
RESULT AND CONCLUSIONThe shapes and sizes of anther and filament were different between the male sterile and the normal plants. For the male sterile plant's, the filament size was no more than 1/2 of that of normal plants and the anthers were shriveled, failed to dehisce and pollinate naturally, and the pollen grains in the anthers had no vitality. Other morphological characteristics were similar between two types of plants. The main reason leading to male sterility of B. chinense was the abnormal development of tapetum cells with two circumstances. The one is that the tapetum cells degraded early during the period of microsporocyte phase to tetrad phase and the other is that the tapetum cells proliferated with delayed degradation in the tetrad to uninucleate phase.
Bupleurum ; anatomy & histology ; cytology ; physiology ; Plant Infertility ; Pollen ; anatomy & histology ; cytology ; physiology
3.Tissue culture of Vinca minor and determination of vincamine.
Entai HOU ; Yingjuan WANG ; Duowei LI ; Na LI ; Hongdong CHEN ; Huaiyu BU
China Journal of Chinese Materia Medica 2011;36(7):823-827
OBJECTIVETo study tissue culture of Vinca minor and determine the content of vincamine.
METHODLeaf blades, stalks, root segment of V. minor were used as explants to study the effect of 2, 4-D,6-BA,NAA on its callus induction and vincamine contents in the orthogonal design experiment. In the peak period of callus formation, vincamine content in callus of V. minor and sterile plants was determined by HPLC. The experimental data was statistically analyzed.
RESULTThe content of 6-BA and NAA had no significant effect on its callus induction. But the content of 2, 4-D had significant effect on its callus induction. Within 20,40,60 d, the content of vincamine in sterile plant was (0.015 +/- 0.003)%, (0.097 +/- 0.001)% , (0.113 +/- 0.06)%, respectively. In the peak period of callus formation, vincamine content in callus of leaf blades, stalks, root segment was (0.024 +/- 0.0025)%, (0.016 +/- 0.0015)%, (0.010 +/- 0.0015)%, respectively. To 30 days of subculture, vincamine content in callus of leaf blades, stalks, root segment was (0.041 +/- 0.002)%, (0.019 +/- 0001)%, (0.016 +/- 0.002)%, respectively.
CONCLUSIONThe optimal hormone combination for callus initiation was MS +2, 4-D 1.0 mg x L(-1) +6-BA 0.5 mg x L(-1) + NAA 0.5 mg x L(-1). In different growth periods, vincamine content in sterile plants is significantly different. From different explants in callus vincamine content is different, in which leaves callus is significantly higher than that of stems, roots produced callus organization.
Culture Techniques ; methods ; Linear Models ; Plant Infertility ; Vinca ; chemistry ; growth & development ; physiology ; Vincamine ; analysis
4.Pollen viability and stigma receptivity of Salvia miltiorrhiza and its relative.
Guocheng ZHONG ; Li ZHANG ; Ruiwu YANG ; Chunbang DING
China Journal of Chinese Materia Medica 2010;35(6):686-689
OBJECTIVETo provide the basal data for artificial cross breeding of Chinese herb Salvia miltiorrhiza from 7 provinces in China and its 4 relatives.
METHODThe pollen viability was evaluated by TTC (2, 3, 5-triphenylte trazolium chloride) test and the stigma receptivity was evaluated by benzidine-H2O2 method.
RESULTThe pollen viability of S. miltiorrhiza from 6 provinces in China and its 4 relatives deceased during time of pollen shedding. Their highest pollen viability was in 2 or 3 days after blooming. But the pollen viability of S. miltiorrhiza (wild and culture) from Hean province in China declined with time after blooming. The most obvious variation of the pollen viability was in S. miltiorrhiza from Shanxi province (RSD 71.3% ) and the least was in wild S. miltiorrhiza from Henan province (RSD 12.4%). The highest average pollen viability was wild S. miltiorrhiza (72.3%) from Henan province while the lowest was S. yunnanensis (38.8%). The stigmas of all the accessions had receptivity when blooming. The stigma receptivity of S. brevilabra was strong in 2 to 4 days after blooming, while the others had less change after blooming. The life span of pollen grains and stigmas could be maintained from 3 to 5 days.
CONCLUSIONThe optimum artificial pollination time of S. miltiorrhiza and its relatives was 2 to 3 days after blooming.
China ; Christianity ; Chromosomes, Plant ; physiology ; DNA, Plant ; analysis ; Flowers ; growth & development ; physiology ; Genetic Variation ; Genetics, Population ; Hydrogen Peroxide ; pharmacology ; Plant Infertility ; physiology ; Plant Proteins ; genetics ; Pollen ; Pollination ; immunology ; physiology ; Polyploidy ; Salvia miltiorrhiza ; physiology
5.Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP.
Qidi ZHU ; Xinbo ZHANG ; M EJAZ ; Gaisheng ZHANG ; Huixue CHE ; Shuping WANG ; Qilu SONG ; Shuling YANG ; Longyu ZHANG
Chinese Journal of Biotechnology 2013;29(5):646-656
Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis
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methods
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Cytoplasm
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metabolism
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DNA, Mitochondrial
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genetics
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DNA, Plant
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genetics
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Gene Expression Profiling
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Genotype
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Plant Infertility
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genetics
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Triticum
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genetics
6.Using single primer PCR to amplify the orf25 gene fragment related to plant male-sterility in Salicornia europaea.
Jin-Yao LI ; Fu-Chun ZHANG ; Ji MA ; Wen-Juan SHAN ; Bin WANG
Chinese Journal of Biotechnology 2003;19(1):120-123
The gene orf25 encodes functional protein that may play an important role in plant fertility control in nature. To clone the orf25 from Salicornia europaea Xinjiang into a T-vector, a single designed primer was used to amplify 1.7kb cDNA fragment with RT-PCR. Sequence analysis reveals that the cloned fragment contains entire orf25 coding region with 98%, 95%, 92% and 88% identity to that of orf25 from Beta vulgaris, Nicotiana, wheat and maize mitochondrion, respectively. This analysis suggests that orf25 gene is highly conserved in terms of evolution in plant; and it also suggests that wild plant Salicornia europaea contain a male-sterility gene similar to crops that is of great importance in improvement of the breed of crop.
Base Sequence
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Chenopodiaceae
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genetics
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DNA Primers
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genetics
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Molecular Sequence Data
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Plant Infertility
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genetics
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physiology
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Plant Proteins
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chemistry
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genetics
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Reverse Transcriptase Polymerase Chain Reaction
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methods
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Sequence Homology, Nucleic Acid
7.The pTA29-barnase chimeric gene transformation of Brassica campestris L. subsp. chinensis Makino var. parachinensis mediated by agrobacterium.
Bihao CAO ; Chengmin MENG ; Jianjun LEI ; Guoju CHEN
Chinese Journal of Biotechnology 2008;24(5):881-886
In order to induce male sterility of Brassica campestris L. subsp. chinensis Makino var. parachinensis, we introduced the chimeric pTA29-barnase gene into it by Agrobacteriumtume faciens transformation. We obtained the transgenic plants, and determined them by PCR, Southern blotting and RT-PCR analysis. Results indicated that the RNase (barnase) gene had been transferred into genome of plant, and its expression level was different among transformation plants. All transgenic plants were male sterile; there was no vigor or a little pollen without fertility in the anther of transgenic plants. The transgenic plants failed to produce seeds under the condition self-control pollination, but hybrid seeds set were obtained when these transgenic plants were cross-pollinated artificially with normal pollen from untransformed plants. Progeny from cross-pollinated maintainer line with transgenic plants segregated in the 1:1 for male sterility and male fertility, and these phenotypes corresponded directly to the presence or absence of the chimeri TA29-barnase gene. The male fertile plants of co-separated progenies could die by spraying 10 mg/L PPT in cotyledon seedling stage. The hybrid F1 between male sterility and other varieties showed heterosis in yield and growth. All these show that it is an efficient method to induce male sterility in Brassica campestris L. subsp. chinensis Makino var. parachinensis by TA29-barnase ene, there is potential on heterosis breeding of Brassica campestris L. subsp. chinensis Makino var. parachinensis.
Agrobacterium tumefaciens
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genetics
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Brassica
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genetics
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growth & development
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Gene Transfer Techniques
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Genes, Plant
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genetics
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Plant Infertility
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genetics
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Plants, Genetically Modified
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genetics
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Ribonucleases
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genetics
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Transformation, Genetic
8.Gene cloning and expression analysis of a male sterility related gene BoDHAR from broccoli.
Guo-Yu ZHANG ; Jun-Gen KANG ; Yan-Guo ZHANG ; Zhi-Hui CHENG ; Xiao-Wu WANG
Chinese Journal of Biotechnology 2006;22(5):751-756
A differentially expressed cDNA fragment obtained from a cDNA-AFLP analysis, which performed on floral buds of male sterile and fertile lines of cabbage, was used as a querying probe to blast the Genbank and Arabidopsis databases. Based on the assembled homologous cDNA sequences, a full-length cDNA of 633 bp for BoDHAR was cloned by RT-PCR. Furthermore, we have experimentally cloned and sequenced the 5' flanking sequence of gene BoDHAR by genomic walking method based on ligation-mediated PCR. The full length DNA sequence with 1486bp, containing two introns, was achieved. Homologous analysis shows that gene has 82.3% identity at nucleotide level, and 79.6% identity at amino acid level with Arabidopsis dehydroascorbate reductase (DHAR) gene AT1 G19570.1. Structurally, BoDHAR encodes a polypeptide of 210 amino acids, which contains a GST-c-DHAR domain highly conserved among other members of the DHAR superfamily and has multiple phosphorylation sites. Promoter predictions software indicated that the 5' upstream region contained putative transcription signals and conserved sequences, one CAAT-box, one G-box and four TGAC-like motifs. To advance our understanding of gene BoDHAR, tissue expression pattern were analyzed by semi-quantitative RT-PCR. The results indicate that expression level of gene BoDHAR is higher in fertile buds than that in sterile buds, and expressed intensively in the anther.
Amino Acid Sequence
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Base Sequence
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Brassica
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genetics
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Cloning, Molecular
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Molecular Sequence Data
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Oxidoreductases
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genetics
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Plant Infertility
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genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Analysis, DNA
9.Effects of Korean red ginseng on semen parameters in male infertility patients: A randomized, placebo-controlled, double-blind clinical study.
Hyun Jun PARK ; Sangmin CHOE ; Nam Cheol PARK
Chinese journal of integrative medicine 2016;22(7):490-495
OBJECTIVETo investigate the effects of Korean red ginseng (KRG) on semen parameters in male infertility patients in a randomized, double-blind, placebo-controlled study.
METHODSA total of 80 male infertility patients with varicocele were recruited from April 2011 to February 2012. The subjects were then divided into the following four groups: non-varicocelectomy (V)+placebo (P) group, V+P group, non-V+KRG group (1.5-g KRG daily), and V+KGR group (1.5-g KRG daily). Semen analysis was performed and hormonal levels were measured in each treatment arm after 12 weeks.
RESULTSAll groups but not the non-V+P group, showed significant improvements in sperm concentrations, motility, morphology, and viability at the end of the study. However, there were no significant differences in serum follicle-stimulating hormone, luteinizing hormone, and testosterone among groups. The incidence of adverse events was low, and all events were assumed to be unrelated to the treatments administered.
CONCLUSIONSAlthough the exact mechanism by which KRG improves spermatogenesis remains unclear, KRG may be a useful agent for the treatment of male infertility. Nevertheless, additional studies to evaluate the optimal dose and duration of treatment are needed.
Adult ; Double-Blind Method ; Hormones ; metabolism ; Humans ; Infertility, Male ; drug therapy ; Male ; Panax ; chemistry ; Placebos ; Plant Extracts ; adverse effects ; pharmacology ; therapeutic use ; Semen ; drug effects ; metabolism
10.Analysis of differential genes and metabolic pathway related to functional male sterility in eggplant.
Zhimin WANG ; Chao YUAN ; Zeqin DING ; Ruolin HU ; Yi NIU ; Qinglin TANG ; Dayong WEI ; Ming SONG ; Yongqing WANG ; Shibing TIAN
Chinese Journal of Biotechnology 2021;37(1):253-265
Based on observing the cytological characteristics of the flower buds of the functional male sterile line (S13) and the fertile line (F142) in eggplant, it was found that the disintegration period of the annular cell clusters in S13 anther was 2 days later than that of F142, and the cells of stomiun tissue and tapetum in F142 disintegrated on the blooming day, while it did not happen in S13. The comparative transcriptomic analysis showed that there were 1 436 differential expression genes (DEGs) (651 up-regulated and 785 down-regulated) in anthers of F142 and S13 at 8, 5 days before flowering and flowering day. The significance analysis of GO enrichment indicated that there were more unigene clusters involved in single cell biological process, metabolism process and cell process, and more catalytic activity and binding function were involved in molecular functions. Through KEGG annotation we found that the common DEGs were mainly enriched in the biosynthesis of secondary metabolites, metabolic pathway, protein processing in endoplasmic reticulum, biosynthesis of amino acids, carbon metabolism and plant hormone signal transduction. The fifteen genes co-expression modules were identified from 16 465 selected genes by weighted gene co-expression network analysis (WGCNA), three of which (Plum2, Royalblue and Bisque4 modules) were highly related to S13 during flower development. KEGG enrichment showed that the specific modules could be enriched in phenylpropanoid biosynthesis, photosynthesis, porphyrin and chlorophyll metabolism, α-linolenic acid metabolism, polysaccharide biosynthesis and metabolism, fatty acid degradation and the mutual transformation of pentose and glucuronic acid. These genes might play important roles during flower development of S13. It provided a reference for further study on the mechanism of anther dehiscence in eggplant.
Flowers/genetics*
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Gene Expression Profiling
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Gene Expression Regulation, Plant
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Humans
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Infertility, Male
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Male
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Metabolic Networks and Pathways/genetics*
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Solanum melongena/genetics*
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Transcriptome/genetics*