1.The physiology of plant seed aging: a review.
Peilin HAN ; Yueming LI ; Zihao LIU ; Wanli ZHOU ; Fan YANG ; Jinghong WANG ; Xiufeng YAN ; Jixiang LIN
Chinese Journal of Biotechnology 2022;38(1):77-88
Seed quality plays an important role in the agricultural and animal husbandry production, the effective utilization of genetic resources, the conservation of biodiversity and the restoration and reconstruction of plant communities. Seed aging is a common physiological phenomenon during storage. It is a natural irreversible process that occurs and develops along with the extension of seed storage time. It is not only related to the growth, yield and quality of seed and seedling establishment, but also has an important effect on the conservation, utilization and development of plant germplasm resources. The physiological mechanisms of seed aging are complex and diverse. Most studies focus on conventional physiological characterization, while systematic and comprehensive in-depth studies are lacking. Here we review the recent advances in understanding the physiology of seed aging process, including the methods of seed aging, the effect of aging on seed germination, and the physiological and molecular mechanisms of seed aging. The change of multiple physiological parameters, including seed vigor, electrical conductivity, malondialdehyde content and storage material in the seed, antioxidant enzyme activity and mitochondrial structure, were summarized. Moreover, insights into the mechanism of seed aging from the aspects of transcriptome, proteome and aging related gene function were summarized. This study may facilitate the research of seed biology and the conservation and utilization of germplasm resources.
Germination
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Plants
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Proteome
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Seedlings
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Seeds/genetics*
2.Karyotypes analysis of Ferula fukanensis.
Xin ZHAO ; Xiao-Jun MA ; Sulaiman KAISAR ; Chang-Liang FU ; Rui-Yang CHEN
China Journal of Chinese Materia Medica 2006;31(2):114-116
OBJECTIVEStudy on the karyotypes analysis of Ferula fukanensis.
METHODThe young roots were treated with 0.000 2 mol x L(1) 8-Hydroxyquinoline for 3 h, carnoy's for 3 h, 1 mol x L(-1) HCl in 5 min,carbol fuchsin coloration for 2 min and the treated roots were utilizied to make the plate for observation.
RESULTThe fukanensis is diploid. The chromosome number of somatic cells was 2n = 22. The karyotype formula is 2n = 2x = 20 = 16m + 4sm. The 4th and 10th are submetacentric, and the others are metacentric.
CONCLUSIONThe karyotype of F. fukanensis belongs to "2A" type of stebbins', and it is a primitive species.
Centromere ; Chromosomes, Plant ; genetics ; Diploidy ; Ferula ; genetics ; Karyotyping ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Seeds ; genetics
3.Species identification of Ligustrum lucidum.
Yu-Shuang WANG ; Yuan-Xi JIN ; Kang-Jia LIU ; Chang GUO ; Yi-Heng WANG ; Chao XU ; Zhi-Xiang ZHANG ; Wen-Pan DONG
China Journal of Chinese Materia Medica 2023;48(11):2940-2948
Ligustrum lucidum is a woody perennial plant of genus Ligustrum in family Oleaceae. Its dried fruit has high medicinal value. In this study, the authors evaluated the variability and species identification efficiency of three specific DAN barcodes(rbcL-accD, ycf1a, ycf1b) and four general DAN barcodes(matK, rbcL, trnH-psbA, ITS2) for a rapid and accurate molecular identification of Ligustrum species. The results revealed that matK, rbcL, trnH-psbA, ITS2 and ycf1a were inefficient for identifying the Ligustrum species, and a large number of insertions and deletions were observed in rbcL-accD sequence, which was thus unsuitable for development as specific barcode. The ycf1b-2 barcode had DNA barcoding gap and high success rate of PCR amplification and DNA sequencing, which was the most suitable DNA barcode for L. lucidum identification and achieved an accurate result. In addition, to optimize the DNA extraction experiment, the authors extracted and analyzed the DNA of the exocarp, mesocarp, endocarp and seed of L. lucidum fruit. It was found that seed was the most effective part for DNA extraction, where DNAs of high concentration and quality were obtained, meeting the needs of species identification. In this study, the experimental method for DNA extraction of L. lucidum was optimized, and the seed was determined as the optimal part for DNA extraction and ycf1b-2 was the specific DNA barcode for L. lucidum identification. This study laid a foundation for the market regulation of L. lucidum.
Ligustrum/genetics*
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Seeds
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Fruit
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Polymerase Chain Reaction
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Research Design
4.Study on seed testing for Salvia miltiorrhiza.
Hong-mei DAN ; Jian-jun QI ; Li-li ZHOU ; Xian-en LI
China Journal of Chinese Materia Medica 2008;33(17):2090-2093
OBJECTIVETo establish a seed testing methods for Salvia miltiorrhiza.
METHODReferring to the International Seed Testing Rules made by ISTA and the Seed Testing for Crops (GB/T3543. 1-1995) issued by China.
RESULTThe seeds are selected by winnowing; the seed purity is about 50%-60%; 100 grain weight is used to determine the quality of the seed; the seed moisture content is determined by air drying, the drying hour is 3 h. Seed viability is tested by TFC method.
Chromosomes, Plant ; genetics ; Germination ; Quality Control ; Salvia miltiorrhiza ; chemistry ; genetics ; physiology ; Seeds ; chemistry ; genetics ; physiology
5.Authenticate of Atractylodes macrocephala seed by amplification refractory mutation system.
Liang CAO ; Chao JIANG ; Hua-Sheng PENG ; Min CHEN ; Yuan YUAN
China Journal of Chinese Materia Medica 2013;38(16):2567-2570
OBJECTIVETo design specific primers and authenticate Atractylodes macrocephala from Atractylodes lancea and A. chinensis.
METHODSNPs in the psbA-trnH sequences of Atractylodes were found by ClustulW program and Bioedit software. Primers for authentic A. macrocephala is designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system.
RESULT172 bp band special for A. macrocephala were found using multi-PCR reaction.
CONCLUSIONThe multi-PCR reaction system could be applied to identify A. macrocephala seed.
Atractylodes ; genetics ; DNA Primers ; genetics ; Mutation ; Polymerase Chain Reaction ; methods ; Quality Control ; Seeds ; genetics
6.Optimization of SRAP & ISSR technology and its application in the identification of seeds of Brassica oleracea L.
Chong LIU ; Cai-Lin GE ; Yun-Ying REN ; Jin-Xiu CHEN ; Xiao-Feng YANG ; Tian-Yue BO
Chinese Journal of Biotechnology 2006;22(4):657-661
In this study, the molecular marker technology of SRAP and ISSR were applied in rapid identification of seeds from eight species of Brassica oleracea L. Firstly, using the genomic DNA of cabbage as template, SRAP and ISSR reaction systems were optimized through testing every factor, respectively, that affects PCR amplification. Then, using the optimized reaction systems, 30 SRAP primer pairs and 15 ISSR primers were applied to amplify genomic DNA of cabbage, savoy, purple cabbage, borecole, cauliflower, broccoli, Brussels sprouts, and kohlrabi The results showed that high polymorphisms were exhibited among the eight species of Brassica oleracea L. by SRAP primer pairs of M3-E5 and M4-E5, as well as ISSR primers of 844 and 888, especially primer 844 which can identify all eight materials efficiently.
Brassica
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genetics
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Nucleic Acid Amplification Techniques
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methods
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Polymorphism, Genetic
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Repetitive Sequences, Nucleic Acid
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Seeds
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genetics
7.Requirements for improved vaccines against foot-and-mouth disease epidemics.
Clinical and Experimental Vaccine Research 2013;2(1):8-18
Inactivated foot-and-mouth disease (FMD) vaccines are currently used worldwide. With the emergence of various FMD virus serotypes and subtypes, vaccines must become more suitable for field-based uses under the current circumstances in terms of the fast and proper selection of vaccine strains, an extended vaccine development period for new viruses, protecting against the risk of virus leakage during vaccine manufacture, counteracting the delayed onset of immune response, counteracting shorter durations of immunity, and the accurate serological differentiation of infected and vaccinated animals and multiple vaccination. The quality of vaccines should then be improved to effectively control FMD outbreaks and minimize the problems that can arise among livestock after vaccinations. Vaccine improvement should be based on using attenuated virus strains with high levels of safety. Moreover, when vaccines are urgently required for newly spread field strains, the seed viruses for new vaccines should be developed for only a short period. Improved vaccines should offer superior immunization to all susceptible animals including cattle and swine. In addition, they should have highly protective effects without persistent infection. In this way, if vaccines are developed using new methods such as reverse genetics or vector vaccine technology, in which live viruses can be easily made by replacing specific protective antigens, even a single vaccination is likely to generate highly protective effects with an extended duration of immunity, and the safety and stability of the vaccines will be assured. We therefore reviewed the current FMD vaccines and their adjuvants, and evaluated if they provide superior immunization to all susceptible animals including cattle and swine.
Animals
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Cattle
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Disease Outbreaks
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Foot-and-Mouth Disease
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Immunization
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Livestock
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Reverse Genetics
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Seeds
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Swine
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Vaccination
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Vaccines
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Viruses
8.Selection of q RT-PCR reference genes for Amomum tsaoko seeds during dormancy release.
Li-Xiang YAO ; Chun-Liu PAN ; Li-Ying YU ; Zhu QIAO ; Mei-Qiong TANG ; Fan WEI
China Journal of Chinese Materia Medica 2021;46(15):3832-3837
Freshly collected seeds of Amomum tsaoko demonstrate obvious dormancy. Therefore, the selection of stable reference genes during seed dormancy release is very important for the subsequent functional research of related genes. In this study, ten commonly used reference genes(GAPDH, 40S, actin, tubulin, EIF4A-9, EIF2α, UBC, UBCE2, 60S, and UBQ) were selected as candidates for quantitative Real-time polymerase chain reaction(qRT-PCR) of the embryo samples of A. tsaoko at different dormancy release stages. Three kinds of software(BestKeeper, geNorm, and Normfinder) and the Delta CT method were used to evaluate the expression stability of the candidate reference genes, and the RefFinder online tool was employed to integrate the results and generate a comprehensive ranking. The results showed that the expression levels of the ten candidate reference genes differed greatly in different embryo samples. GAPDH and UBC had high expression levels, as manifested by the small Ct values. GeNorm identified 40S and UBCE2 as the most stable genes. NormFinder ranked EIF2α as the most stable gene and UBC as the least stable gene. UBCE2 was found to be the most stable gene and actin the least stable one by BestKeeper. Delta CT analysis suggested that the expression of 40S was most stable. UBCE2 was recommended as the most stably expressed gene by RefFinder. Thus, UBCE2 is the ideal reference gene for qRT-PCR analysis of A. tsaoko seeds at different dormancy release stages. The results may lay a foundation for analyzing the expression of related genes during seed dormancy release of A. tsaoko.
Amomum
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Gene Expression Profiling
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Real-Time Polymerase Chain Reaction
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Reverse Transcriptase Polymerase Chain Reaction
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Seeds/genetics*
9.Primary study on measuring the internal transcribed spacer I regions of rRNA genein seeds of Gentiana dahurica.
Ke-ping JI ; Xi-ling ZHANG ; Li-sha LIU ; Quan-yun LU ; Che CHENG
China Journal of Chinese Materia Medica 2003;28(4):313-316
OBJECTIVETo amplify the PCR with the internal transcribed spacerl regions measure the base sequence of the amplified products of DNA, and to set up an identified standard on the level of molecule.
METHODDNA from the seeds of G. dahurica was extracted by conventional method, and composed peculiar primer was used to amplify with the internal transcribed spacerl regions of the rRNA gene, and the base sequence of the amplified products by stopping the circle of the end of double deoxidation of four color fluorescent mark was measured.
RESULTIt was proved by agar sugar gel electrophoresis that the PCR amplified products of the internal transcribed spacerl regions of the rRNA gene existed. The base sequence of the seeds of G. dahurica's internal transcribed spacerl regions of the rRNA gene was measured.
CONCLUSIONTo measure the base sequence of internal transcribed spacerl regions of the rRNA gene in the seeds of G. dahurica's is a method to identify vegetal Chinese traditional medicine on the level of molecule.
Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Gentiana ; genetics ; Molecular Sequence Data ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; Seeds ; genetics
10.Callus induction and regeneration from mature seeds of indica rice minghui 63 and anti-fungal assay of transgenic rice plants.
Li-Jiang WANG ; Xiao-Tian MING ; Cheng-Cai AN ; Hua-Yi YUAN ; Zhang-Liang CHEN
Chinese Journal of Biotechnology 2002;18(3):323-326
A large number of callus from mature seeds of indica rice minghui 63 were obtained through pre-induction on medium with 2 mg/L 2,4-D but without inorganic and organic components for 9 days. Trichosanthin gene was transferred into indica rice minghui 63 by using agrobacterium with the help of bombardment and the transgenic plants were obtained by inducing regeneration. Southern and Western blot analysis showed that the trichosanthin gene had been transferred into genome of minghui 63 and expressed in rice plants. The anti-fungal assay suggested that transgenic rice plants enhanced resistance to infection of Pyricularia oryzae.
Mitosporic Fungi
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drug effects
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Oryza
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genetics
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microbiology
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Plant Diseases
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microbiology
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Plants, Genetically Modified
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Regeneration
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Seeds
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physiology
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Trichosanthin
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genetics