1.Advances in identification methods of alien genomic components in plants.
Zhongyi XIE ; Jiangbo DANG ; Guo WEN ; Haiyan WANG ; Qigao GUO ; Guolu LIANG
Chinese Journal of Biotechnology 2021;37(8):2703-2718
Plants with alien genomic components (alien chromosomes / chromosomal fragments / genes) are important materials for genomic research and crop improvement. To date, four strategies based on trait observation, chromosome analysis, specific proteins, and DNA sequences have been developed for the identification of alien genomic components. Among them, DNA sequence-based molecular markers are mainly used to identify alien genomic components. This review summarized several molecular markers for identification of alien genomic components in wheat, cabbage and other important crops. We also compared the characteristics of nine common molecular markers, such as simple sequence repeat (SSR), insertion-deletion (InDel) and single nucleotide polymorphism (SNP). In general, the accuracy of using a combination of different identification methods is higher than using a single identification method. We analyzed the application of different combination of identification methods, and provided the best combination for wheat, brassica and other crops. High-throughput detection can be easily achieved by using the new generation molecular markers such as InDel and SNP, which can be used to determine the precise localization of alien introgression genes. To increase the identification efficiency, other new identification methods, such as microarray comparative genomic hybridization (array-CGH) and suppression subtractive hybridization (SSH), may also be included.
Chromosomes, Plant
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Comparative Genomic Hybridization
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Genome, Plant/genetics*
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Genomics
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Triticum/genetics*
2.Molecular and cytogenetic identification of Triticum aestivum-Leymus racemosus translocation line T6DL·7LrS.
Chinese Journal of Biotechnology 2018;34(11):1823-1830
Leymus racemosus had a high resistant capacity to wheat scab (Fusarum head blight). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the germplasm of wheat resistance. To obtain Triticum aestivum-Leymus racemosus translocation line with scab resistance, we irradiated the pollen of T. aestivum-L. racemosus disomic addition line DA7Lr by ⁶⁰Co-γ-rays 1 200 R (100 R/min) prior to pollinating to emasculation T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 by GISH. The plant with one translocation chromosome was self-pollinated, and at meiotic metaphase I its progenies with two translocation chromosomes were analyzed for chromosome pairing behavior in their pollen mother cells (PMCs). One rod bivalent was observed at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. Sequential GISH-FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line was confirmed as T6DL·7LrS. The translocation line had higher resistance to wheat scab and feasibility to be used as a new source in wheat breeding resistant to scab disease.
Chromosomes, Plant
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Disease Resistance
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genetics
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In Situ Hybridization, Fluorescence
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Plant Breeding
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Plant Diseases
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genetics
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Poaceae
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genetics
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Pollen
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Translocation, Genetic
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Triticum
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genetics
3.Optinization of rapid propagation technique and induction and identification of autotetraploid of Polygonum multiflorum.
He-Ping HUANG ; Shan-Lin GAO ; Jian WANG ; Lu-Qi HUANG ; Peng HUANG
China Journal of Chinese Materia Medica 2013;38(10):1467-1470
OBJECTIVETo establish and optimize the rapid propagation system of Polygonum multiflorum, as well as explore method for induction and identification of autotetraploid.
METHODPropagation medium was optimized by orthogonal test. The buds were immersed in colchicine solution with different concentrations for different time to select induction conditions for autotetraploid of P. multiflorum.
RESULTThe most appropriate propagation medium was MS medium supplemented with 1.0 mg x L(-1) 6-BA, 0.3 mg x L(-1) NAA, and 0.4 mg x L(-1) PP333. That the buds were soaked in 0.2% colchicine solution for 30 h, or soaked in 0.3% colchicine solution for 18 h, was optimal condition to induce autopolyploid of P. multiflorum with induction rate as high as 16.7%.
CONCLUSIONRapid propagation of P. multiflorum could be achieved by tissue culture. Furthermore, colchicine was an effective inducer of polyploidy, and 25 tetraploid lines were obtained through chromosome identification. The experiment laid a foundation for the wild resource conservation, superior varieties breeding of P. multiflorum.
Chromosomes, Plant ; genetics ; Culture Media ; metabolism ; Polygonum ; genetics ; growth & development ; metabolism ; Tetraploidy ; Tissue Culture Techniques ; methods
4.Production and characterization of a somatic hybrid of Chinese cabbage and cabbage.
Chinese Journal of Biotechnology 2012;28(9):1080-1092
In order to broaden Chinese cabbage gene pool, we conducted interspecific somatic hybridization between Chinese cabbage (Brassica campestris, 2n=20, AA) and Cabbage (B. oleracea, 2n=18, CC). Protoplasts were isolated from 10-day-old cotyledons and hypocotyls of young seedlings, and fused by 40% polyethylene glycol (PEG). Fused cells were cultured in modified K8p liquid medium supplemented with some plant growth regulators. Fusion products were characterized by their morphological, cytological and molecular biological traits. The results showed that, a total of 35 regenerated green plants were obtained from 320 calli, the plant regeneration frequency was 10.94%, and eleven of which were survived in greenhouse. All regenerants were true hybrids as confirmed by randomly amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) analysis. Ploidy levels of hybrid plants were determined by chromosome counting and flow cytometry. The sum of the chromosome number (2n = 38) from the two fusion patents were found in 36.4% of regeneratns; another 36.4% had chromosomes range to 58-60; 27.2% had more chromosomes ranges to 70-76. All regenerated plants produced normal flowers. We investigated the pollen fertility and seed set after self-pollination and backcrossing with the parental species. For hybrids with chromosomes more than 38 it was possible to obtain some seeds when they after self-pollination. Within the group of hybrids with 38 chromosomes, seed set were very variable, only 0.11 seeds per pod by self-pollination, 0.23-0.76 by open-pollination, 0.02-0.04 by backcrossing with Chinese cabbage. Progeny lines obtained by self-pollination had larger leaves and leaf shapes intermediate of the parental species. Pollen fertility was gradually recovered in the first and second progenies. The backcrossing progeny lines, as a whole, exhibited morphologies were similar to Chinese cabbage. Morphological variations were observed among the somatic hybrids and their progenies.
Brassica napus
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genetics
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growth & development
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Breeding
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Chromosomes, Plant
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Hybridization, Genetic
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genetics
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Mustard Plant
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genetics
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growth & development
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Plant Somatic Embryogenesis Techniques
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Ploidies
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Pollen
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physiology
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Protoplasts
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cytology
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Random Amplified Polymorphic DNA Technique
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Recombination, Genetic
5.Karyomorphology research in seven kinds of dandelion in Northeast.
Wei NING ; Jie WU ; Ting ZHAO ; Xin ZHAO ; Tianlai LI
China Journal of Chinese Materia Medica 2012;37(6):771-776
OBJECTIVETo determine cytology chromosome numbers, doubling, type, karyotype formulae of seven kinds of Taraxacum.
METHODSeven kinds of dandelion in Northeast China were karyomorphologically studied by conventional pressed slice method.
RESULTThe interphase nuclei and prophase chromosomes of all species were found to be of the complex chromocenter type and the interstitial type respectively. The somatic chromosomes showed polysomaty within or among species. The range of chromosome numbers was from 16 to 32, and 24 was preponderant. It is firstly confirmed that the chromosome numbers of T. ohwianum were 2n = 2x = 16 of and those of T. variegatum 2n = 4x = 32 of, and those of other five kinds of dandelion 2n = 3x = 24. Karyotype was diverse and consisted of metacentric, submetacentric and satellites. The relative lengh of chromosome varies from 3.74 to 27.68, asymmetry index was between 59.68% and 64.02%; The karyotype type was belonged to "1A", "2A"and "2B" extensively, to ensure the T. ohwianum and T. variegatum evolutional karyotype type and genetic inheritance pattern.
CONCLUSIONAccording to results of the cytology karyotype we suggest that T. antungense and T. urbanum could be merged, the result was consistent with Flora Reipublicae Popularis Sinicae (FRPS). This paper reports systematically cytology karyotype feature of seven kinds of dandelions in Northeast, provides the cytology theoretical basis for further development and use of the resource and genetic breeding research of dandelions.
China ; Chromosomes, Plant ; genetics ; Karyotype ; Plants, Medicinal ; classification ; cytology ; genetics ; Taraxacum ; classification ; cytology ; genetics
6.Karyotype analysis and its systematic implications of Pterocypsela formosana and P. elata.
Daigui ZHANG ; Xiaoqi XIANG ; Jieying ZHU ; Gongxi CHEN ; Tao DENG ; Yongxin CHEN
China Journal of Chinese Materia Medica 2012;37(11):1527-1531
Pterocypsela is a very important traditional Chinese medicine from the tribe Cichorieae of Asteraceae. Mitotic chromosome numbers and karyotypes are reported for P. formosana and P. elata from Hunan and Hubei province, China. The former is new and the latter provide confirmation of previous reference. All P. taxa are diploidy with 2n = 18 and their basic number is tentatively suggested as x = 9. Karyotype of Pterocypsela is 2A and P. formosana with a karyotype formula of 2n = 2x = 18 = 4m + 14sm, and 2n = 2x = 18 = 2m + 8sm +8st for P. elata. It is the first time to report the AI value for Pterocypsela in this paper. Cytological data of chromosomal numbers and karyotypes were used to discuss the close relationships of the Pterocypsela genus and the taxonomy of the medicinal plants.
Asteraceae
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classification
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cytology
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genetics
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Chromosomes, Plant
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genetics
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Diploidy
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Karyotyping
7.Tissue culture and polyploidy induction of Morinda officinalis.
Meizhen LIN ; Qingzhen WU ; Song ZHENG ; Huiqiao TIAN
China Journal of Chinese Materia Medica 2011;36(17):2325-2328
OBJECTIVETo establish an effective protocol for plant generation and induce polyploidy of Morinda offcinalis.
METHODCallus was induced from immature embryo of M. officinalis and polyploidy was inducted by using colchicine treatment method. Chromosome was detected by flow cytometry.
RESULT AND CONCLUSIONThe highest induction rate of polyploidy was 18.40%, which was obtained with 500 mg x L(-1) colchicine treatment for 5 days. Roots of polyploid were bigger than diploid. Advantages of using immature embryo as explants are easy for sterilization, higher rate of callus induction and low degree dedifferentiation. The induced polyploidy of M. officinalis may have a value for spread of cultivation.
Chromosomes, Plant ; genetics ; Morinda ; genetics ; growth & development ; Polyploidy ; Tissue Culture Techniques ; methods
8.Chromosome characteristics of three Coptis species.
Gang XIANG ; Tihua FU ; Qiaojia FAN
China Journal of Chinese Materia Medica 2010;35(11):1367-1371
In the present paper, three Coptis species, collected from Sichuan and Chongqing, China, were used for karyotypic analyses. The results indicated that both C. chinensis and C. omeinensis were diploid with chromosome 2n = 2x = 18, and C. deltoidea was an autotriploid with chromosomes 2n = 3x = 27, which explained why this species was morphologically so isolated from other species and its sterile and narrow distributing regions. The relationship between C. chinensis and C. omeinensis based on chromosome data was discussed. The probable origin of C. deltoidea was also suggested.
China
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Chromosomes, Plant
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genetics
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Coptis
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genetics
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Diploidy
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Karyotyping
9.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
10.Staining and slide-preparing technique of mitotic chromosomes and application in karyotype determination of Ephedra.
Jiling WU ; Sheng LI ; Hanyu JIANG ; Yuhong GAO ; Junyi NIU
China Journal of Chinese Materia Medica 2009;34(21):2725-2729
OBJECTIVETo study the karyotype of four Ephedra plants in order to provide the cytologic evidence for the genetic diversity and identification genetic resources of Ephedra.
METHODThe roots of germinating seeds were used to study the karyotype of four Ephedra plants by staining and slide-preparing technique of mitotic chromosomes.
RESULTthe optimal root-sampling time was about 10: 20 - 10:40 am. Using 0.002 mol x L(-1) 8-Hydroxyquinoline to pretreating the intravital root tips, the optimal pretreatment time for E. Sinica, E. intermedina, E. equisetina and E. przewalskii was 4, 5, 4.5 and 3.5 h, respectively. E. przewalskii and E. equisetina were diploid, E. Sinica and E. intermedina were belonged quadruple. The karyotype formulae of the four species were 2n = 2x = 14 = 2M + 8m + 4sm, 2n = 2x = 14 = 10m + 4st, 2n = 4x = 28 = 20m (2SAT) +8st, and 2n = 4x = 28 = 20m (SAT) + 6st + 2sm, respectively.
CONCLUSIONAll the karyotypes of four Ephedra species were 2A type, which was the symmetric karyotype.
Chromosomes, Plant ; genetics ; Ephedra ; cytology ; genetics ; Karyotyping ; methods ; Mitosis

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