1.Screening of fluorescent bacteria for growth promotion and biocontrol potential against Pyricularia oryzae on aerobic rice (MARDI Aerob 1)
Ng Lee Chuen ; Nur Atiqah Maznan ; Norhayati Yusuf ; Andrew A. Ngadin
Malaysian Journal of Microbiology 2021;17(1):52-59
Aims:
This study aimed to screen the plant growth-promoting fluorescent bacteria (FLB) which isolated from the healthy
rice rhizophere and to evaluate its biocontrol and growth promotion properties against Pyricularia oryzae on aerobic rice
seedling of MARDI Aerob 1.
Methodology and results:
King’s B agar with glycerol was used as the selective medium to isolate FLB from the
healthy rice rhizosphere soil. All FLB obtained were in vitro screened for antagonistic activities against P. oryzae using
dual culture, volatile substances and hydrogen cyanide productions. The potential FLB isolates were further evaluated
on rice seedling early growth promotion before identified using 16S rRNA gene sequencing. A total of 24 FLB were
isolated from the healthy rice rhizosphere soil in Setiu, Terengganu, Malaysia. Isolates: FLB4, FLB5, FLB7 and FLB10
scored the total of percentage inhibition radial growth (PIRG) values ranged 99.5-105.0%. Further seedling growth
promotion screening revealed that FLB4, FLB7 and FLB10 were significantly improved seedling growth with vigor index
of 378.32%, 461.53% and 335.60% over control (133.31%). 16S rRNA sequencing identified that FLB7 as Bacillus
subtilis and the FLB4 and FLB10 as Pseudomonas putida.
Conclusion, significance and impact of study
The selected FLB isolates (FLB4, FLB7 and FLB10) are potential to be
developed as biological control agents against P. oryzae with growth promoting property on aerobic rice seedling.
Plant Breeding--methods
2.Isolation and identification of pathogen causing damping off at seedling stage of Trollius chinensis.
Yong LI ; Rong WANG ; Ruo-Fan WEI ; Wan-Long DING
China Journal of Chinese Materia Medica 2021;46(12):3102-3105
Trollius chinensis is a traditional Chinese medicinal material in China, the wild resource of T. chinensis are now exhausted, and commercial medicinal T. chinensis mainly depends on artificial cultivation. As one of the most severely happened diseases at the seedling period, damping off has been a serious threaten to the breeding of T. chinensis seedlings. However, no related research have been reported so far. So, the authors collected damping-off samples of T. chinensis in 2018 from seedling breeding nursery in Guyuan, Hebei province, and carried out study on taxonomic identification of the pathogen. Damping off occurs in the T. chinensis production area from mid-May to late June every year. At the beginning, brown lesions were observed on the basal stem, then the lesions circumferential expanded and constricted, and finally resulted in the fall and death of T. chinensis seedlings. Pathogenic isolate was growing rapidly on the PDA medium, well developed aerial mycelia were grey white at first, then turned brown gradually, and a great number of small dark brown sclerotia were developed in the middle and periphery of the colony. Mycelial diameter of the pathogen was about 7 to 10 μm, near right angle or acute angle branches, near branches with septa, branches and septa with constriction. After the healthy T. chinensis seedlings were inoculated by pathogenic isolate, damping-off was observed soon, and the symptom was as same as those observed in the field. Through homogenous blast, the rDNA-ITS sequence of the pathogenic isolate shown 99.49% to 99.84% homology with Rhizoctonia solani, R. solani AG-1 IC mycelium anastomosis group and Thanatephorus cucumeris, the sexual type of Rhizoctonia. Furthermore, obvious mycelial anastomosis phenomena were observed when the pathogenic isolate and R. solani AG-1 IC strain were confronting cultured. Based on the results above, the pathogenic isolate causing damping off of T. chinensis was identified as R. solani AG-1 IC mycelial anastomosis group. RESULTS:: in the present work have important significance for further research on basic biology of the pathogen and integrated control of damping off causing by it on T. chinensis.
Basidiomycota
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Plant Breeding
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Plant Diseases
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Rhizoctonia
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Seedlings
3.Breeding peanut variety Yuhua 7 by fast neutron irradiation and tissue culture.
Xia WANG ; Luxiang LIU ; Lixian QIAO ; Jiongming SUI ; Defeng JIANG ; Guan LI ; Linshu ZHAO ; Jingshan WANG
Chinese Journal of Biotechnology 2019;35(2):270-280
Creating new germplasms and breeding new cultivars in peanut by radiation mutagenesis and tissue culture were conducted in this study, aiming to develop new breeding method of peanut. Mature seeds from Luhua 11, the most commonly grown peanut cultivar in Northern China, were treated by fast neutron irradiation. Then the embryo leaflets were separated from the irradiated seeds and inoculated on the media, and the regenerated seedlings were obtained through somatic embryogenesis pathway. The regenerated seedlings were grafted, acclimated and then transplanted into field and the selfed pods were harvested from 83 regenerated plants. The progenies were selected by the pedigree method, and 107 mutants were obtained from the progenies of the 83 regenerated plants. Different mutants showed obvious variation in many agronomic traits, including main stem height, branch number, pod shape and size, seed coat color, inner seed coat color, oil content and protein content etc. Yuhua 7, a new peanut variety with low oil content, early maturity and waterlogging tolerance was obtained. The yield of Yuhua 7 was over 14% higher than that of the mutagenic parent Luhua 11, and the oil content of kernels was 47.0%, lower than that of parent Luhua 11 with 52.1% oil content. Yuhua 7 had passed peanut variety regional multi-location trials in Liaoning Province in 2016 and its average yield was 13.8% higher than that of the control variety Baisha 1017. It had also passed national peanut variety registration, and the registration ID is "GPD peanut (2018) 370105". The results show that irradiation mutagenesis combined with tissue culture is an effective method for creating new germplasm and breeding new varieties of peanut.
Arachis
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Breeding
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China
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Fast Neutrons
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Plant Breeding
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Seeds
4.Development direction of molecular breeding of medicinal plants.
Wen-Guang WU ; Lin-Lin DONG ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2020;45(11):2714-2719
To breed new varieties of medicinal plants with high resistance is the premise to ensure the production of high-quality medicinal materials. Molecular breeding using modern molecular biology and genetic technology can save time and effort and realize rapid and accurate breeding. Here we are trying to summarize the difference of breeding characteristics between medicinal plants and crops such as genetic background and breeding purpose. The strategy of molecular breeding of medicinal plants was summarized, and the four-phases breeding based on high-throughput sequencing and target gene mining was emphasized. We put forward the current molecular breeding of medicinal plants in the condition of four phases breeding is the optimal technological way of breeding, and gene editing breeding is the direction of medicinal plants breeding.
Breeding
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DNA Shuffling
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Gene Editing
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Plant Breeding
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Plants, Medicinal
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genetics
5.Breeding on a new peanut variety Yuhua91 with high oleic acid content.
Leilei PAN ; Yanan JIANG ; Wenjie ZHOU ; Pingping JIANG ; Lanrong WU ; Ao CHEN ; Hong ZHU ; Jiongming SUI ; Jingshan WANG ; Lixian QIAO
Chinese Journal of Biotechnology 2019;35(9):1698-1706
Yuhua91 is a new peanut variety with high oleic acid content bred by Qingdao Agricultural University. The crossing was conducted with Luhua11 as female parent and with Kainong1715, an F435-type variety with high oleic acid content as male parent. The real F1 hybrids were screened by sequencing on PCR amplification products, and those homozygotes with bb genotype in F2 populations were screened by the same sequencing method as above. The content of oleic and linoleic acid was measured on the kernels harvested from F2 single plants by near infrared ray method, and those kernels whose content of oleic was above 80%, oleic and linoleic acid ratio was above 10.0 were obtained and planted into a row, with pedigree method for subsequent selection breeding. Yuhua91 has some characters of small pod, light and obvious pod texture, 148.06 g per 100 pods, 63.31 g per 100 kernels, 75.15% shelling percentage, long elliptic seed kernel, pink seed coat, without crack, white endotesta. Its content of protein, oil, oleic acid, linoleic acid and palmitic acid was 26.57%, 52.72%, 80.40%, 2.50% and 5.57% respectively. Yuhua91 has other characters of strong seedlings, compact pod areas, and moderate resistance to leaf spot disease and bacterial wilt. Average pod yield is 215.79 kg per Mu, 15.27% higher than the control variety Huayu20. Average seed kernels yield is 157.33 kg per Mu, 21.64% higher than the control variety Huayu20. Yuhua 91 has been registered on department of agriculture in 2018, and the registration No. is GPD peanut (2018) 370210, fit for growing in Shandong Province.
Arachis
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Oleic Acid
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Plant Breeding
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Seeds
6.Directional breeding of high oil content peanut variety Yuhua 9 by in vitro mutagenesis and screening.
Jingshan WANG ; Yanan JIANG ; Xiubo YIN ; Yanjun YI ; Jian ZHAO ; Puxiang SHI ; Songjian LI ; Shanlin YU
Chinese Journal of Biotechnology 2019;35(7):1277-1285
Leaf water potential of peanut subjected to drought stress is positively related to the oil content of peanut kernels. The aim of this study was to directly screen the high oil mutants of peanut and create the new peanut varieties using hydroxyproline as water potential regulator. In vitro mutagenesis was carried out with the embryonic leaflets of peanut variety Huayu 20 as explants and pingyangmycin as a mutagen added into the somatic embryo formation medium. The formed somatic embryos were successively transferred to somatic embryo germination and selection medium containing 6 mmol/L hydroxyproline (at -2.079 MPa water potential ) to induce regeneration and directionally screen high oil content mutants. After that, these plantlets were grafted and transplanted to the experimental field and 132 high oil mutants with oil content over 55% were obtained from the offspring of regenerated plants. Finally, among them, the oil contents of 27 lines were higher than 58% and of 2 lines were higher than 60%. A new peanut variety Yuhua 9 with high yield and oil content was bred from the regenerated plant progenies combining the pedigree breeding method. The yield was 14.0% higher than that of the control cultivar in the testing new peanut varieties of Liaoning province, and also it has passed the national registration of non-major crop varieties. Yuhua 9 with an oil content of 61.05%, which was 11.55 percentage points higher than that of the parent Huayu 20, was the peanut cultivar with the highest oil content in the world. The result showed that it was an effective way for directional breeding of high oil peanut varieties by means of the three-step technique including in vitro mutagenesis, directional screening by reducing water potential in medium and pedigree selection of regenerated plant progenies.
Arachis
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Droughts
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Germination
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Mutagenesis
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Plant Breeding
7.Progress on germplasm enhancement and breeding of Rehmannia glutinosa.
Feng-Qing WANG ; Cai-Xia XIE ; Rui-Bin SUN ; Zhong-Yi ZHANG
China Journal of Chinese Materia Medica 2018;43(21):4203-4209
The history of Rehmannia glutinosa breeding has already beyond 100 years. There are rich cultivated varieties and wild germplasm resources in R. glutinosa. However, there also exist a lot of problems, such as, the pedigree of the existing varieties is not clear, the genetic basis is narrow, backward method of germplasm enhancement and breeding. Breeding of new varieties has been unable to meet the demand of R. glutinosa production in the new era. This paper summarizes the species of Rehmannia and their distribution, the diversity of plant morphology and the quality of R. glutinosa germplasm resources, as well as the progress of R. glutinosa breeding in recent 100 years. For ensuring the orderly, effective and safe production of R. glutinosa, the authors suggest to establish the wild resources protection area and germplasm resources garden, deeply study the genetic base of quality, strengthen application of new breeding method such as mutation breeding, haploid breeding and gene editing.
Plant Breeding
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Plants, Medicinal
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genetics
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Rehmannia
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genetics
8.Research progress of tissue culture of Aconitum.
Bi-Wei YIN ; Liu-Ying MAO ; Ling-Li CHEN ; Mei TIAN ; Guang-Hong CUI
China Journal of Chinese Materia Medica 2020;45(23):5645-5649
Aconitum is a kind of important medicinal plant, which has been used in China for more than 2 000 years, with both a good medicinal and ornamental value. However, due to the lack of effective breeding methods and low seed and root propagation coefficients, the comprehensive development and utilization of Aconitum were greatly restricted. Tissue culture is an important basis for seed selection, germplasm conservation and genetic engineering. Therefore, this paper summarized the research on tissue culture of Aconitum, put forward the main problems and corresponding countermeasures, and provided important references for accelerating the seedling breeding of Aconitum and conducting the basic research of molecular biology.
Aconitum
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China
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Plant Breeding
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Plants, Medicinal
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Seedlings
9.Investigation on reproductive characteristics of Polygonatum cyrtonema.
Li-Ge LI ; Ze-Rui ZHANG ; Yan SHI ; Jing-Jing LIU ; Ying-Xiong QIU ; Dong-Hong CHEN ; Jin-Ping SI
China Journal of Chinese Materia Medica 2021;46(5):1079-1083
The study is aimed to investigate the reproductive biology characteristics of Polygonatum cyrtonema, especially including phenology, flower bud differentiation, flowering timing, floral traits, pollen vigor and stigma receptivity. The results showed that P. cyrtonema forms inflorescence before the leaves spread. In the wild, P. cyrtonema is mainly pollinated by insects such as bumblebees, with a seed setting rate of 65.12%. The seed setting rate of indoor single plant isolation or self-pollination enclosed by parchment paper bag is 0, indicating that it is self-incompatible. In Lin'an city, seedlings begin to emerge from mid-March to early April(the temperature is higher than 7.5 ℃), buds begin to emerge from the end of March to mid-April, and then undergo the full bloom stage from mid-to-late April, and the final flowering stage from the end of April to mid-May. The whole flowering period lasts 36 to 45 days. There are obvious differences in the phenology of different provenances. The flowers come into bloom from the base to the top along the aboveground main axis, which usually contain 4-22 inflorescences with(2-) 4-10(-21) flowers per inflorescence. The flowering pe-riod for a single plant is 26-38 days. The single flower lasts about 20-25 days from budding to opening and withers 2 days after pollination, and then the ovary will gradually expand. If unpollinated, it will continue to bloom for 3-5 days and then wither. Flower development period is significantly related to pollen vigor and stigma remittance. The pollen viability is the highest when the flower is fully opened with anthers gathering on the stigma, and the receptivity is the strongest when the stigma protrudes out of the perianth and secretes mucus. The fruits and seeds ripen in October, and proper shading can ensure the smooth development and maturity of the seeds. This study provides a basis for the hybrid breeding and seed production of P. cyrtonema.
Flowers
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Plant Breeding
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Pollination
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Polygonatum
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Reproduction
10.Chloroplast genome in Malus floribunda Siebold.
Xun WANG ; Ziquan FENG ; Daru WANG ; Yuepeng HAN ; Xiaofei WANG ; Xiang SHEN ; Chunxiang YOU
Chinese Journal of Biotechnology 2022;38(10):3713-3727
Malus floribunda Siebold. (Malus) is widely cultivated all over the world, which is of high ornamental value and breeding significance. Comparative analysis of the chloroplast genome can help enrich the phylogenetic relationship and facilitate germplasm utilization of Malus. Based on the whole genome sequencing data, a complete chloroplast genome (M. floribunda) with tetrad structure was assembled. The chloroplast genome (160 037 bp) was composed of a large single-copy (LSC) region (88 142 bp), inverted repeat (IR) B (26 353 bp), a small single-copy (SSC) region (19 189 bp), and IRA (26 353 bp). A total of 111 genes were annotated: 78 protein-coding genes, 29 tRNA genes and 4 rRNA genes. In addition, a large number of repeat sequences were identified in the genome, which was slightly different from that of M. sieboldii and M. toringoides. As for the relative synonymous codon usage, 30 high-frequency codons were found, and the codons tended to end with A/T. The results of interspecific sequence alignment and boundary analysis suggested the sequence variation of the LSC region was large, and the expansion and contraction of the SC region and IR region of the eight Malus species were generally similar. According to the phylogenetic analysis of chloroplast genome sequences, M. floribunda, M. hupehensis, and M. toringoides were grouped into one clade. The findings in this study can provide data support for the development of genetic markers and utilization of germplasm resources in the future.
Genome, Chloroplast
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Malus
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Phylogeny
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Plant Breeding
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Codon