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Application and Prospect of Biochar and Biochar-Based Fertilizer in Tobacco Agriculture
WANG Chengji, TANG Lina, HU Zhongliang, LI Yanchun, HUANG Yibin
2021, 35 (4): 997-1007. DOI: 10.11869/j.issn.100-8551.2021.04.0997
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Tobacco is an important economic crop in our country. Improving the quality and value of tobacco leaf and implementing green and clean production of tobacco are the inevitable trend of green development of tobacco agriculture. It is a win-win approach to realize the resource utilization and environmental friendly development from tobacco stalk carbonization and returning to the field. Biochar has a large specific surface area, developed pore structure, abundant functional groups, stable ring structure and good adsorption properties, which give it great potential to improve continuous cropping obstacle tobacco soil and rebuild healthy microbial habitat. Biochar-based fertilizer combines the advantages of biochar and fertilizer, and has the peculiarities over its raw material. It can slowly release nutrients, maintain fertilizer efficiency for a long time, continuously improve soil environment, promote the healthy growth of tobacco, improve the quality and value of tobacco leaf, and has great potential in tobacco agriculture. This paper reviewed the effects of biochar and biochar-based fertilizer application on tobacco-planting soil physical and chemical properties, soil nutrient, greenhouse gas emissions, soil microorganism and enzyme activity, flue-cured tobacco growth and quality of tobacco leaves, tobacco soil-borne disease. The problems existing in the utilization of biochar and biochar-based fertilizer in tobaocco agriculture were analyzed objectively and then put forward the future research direction: (1) The mechanism of biochar and biochar-based fertilizer regulating the quality of tobacco-planting soil and the growth and quality of tobacco. (2) Ecological and environmental risk assessment of biochar and biochar-based fertilizer application in tobacco-planting soil. (3) Innovation in production technology and process of biochar and biochar-based fertilizer. The results can provide scientific basis for the rational application of biochar and biochar-based fertilizer in tobacco agriculture.
Screening of Imidazolinone-Resistant Rice From EMS-Mutated Populations
CHEN Tianzi, YU Yue, LING Xitie, ZHANG Baolong
2021, 35 (2): 253-261. DOI: 10.11869/j.issn.100-8551.2021.02.0253
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In order to create non-transgenic herbicide-tolerant rice germplasm which would control the weed easily in rice simplified cultivation in China, rice seeds were treated with 0.5% (w/v) ethyl methanesulfonate for 14 h. The resulted M 2 generation at 3-4 leaf stage were sprayed with 160 mg·L -1 of imazapic. The non-resistant seedlings turned into yellow, stunted or even died, while the resistant seedlings were normal and the plant height increased evidently. The resistance of M 2 to imazapic was stably inherited to their progeny M 3, reached up to 1 200 mg·L -1 imazapic, and 100 times as high as the wild type did. The acetolactate synthase( ALS) genes in the imidazolinone-resistant M 3 mutants and the wild-type were cloned by PCR. Sequence analysis revealed that the open reading frame of ALS gene in the imidazolidone-resistant M 3 plants carry two points mutations A1879C and G1880T, which results in a substitution of serine(AGT) to leucine(CTT) at amino acid 627, or a point mutation G1880A, which results in a substitution of serine(AGT) to asparagine(AAT) at amino acid 627. A colorimetry method was used to compare the enzyme sensitivity of ALS to imazapic between the herbicide resistant M 3 mutant and the wild type. The tolerance of ALS enzyme to imazapic in the imidazolinone resistant rice was 109-fold of that in the wild type. Taken together, imidazolidone- resistant rice mutants with substitution of serine to leucine at amino acid 627 was a novel mutation and was first reported. The imidazolidone-resistant rice obtained in this study showed stable and high resistance to imazapic, make it potential in the application of weed control.
Analysis of Small RNA Sequencing Variation in Rice Irradiated by 60Co-γ Ray
ZHANG Li, ZHANG Yong, LEI Xingyu, ZHANG Yuanhai, ZHANG Yiyan, PENG Xuanming, YANG Zhen
2021, 35 (11): 2441-2450. DOI: 10.11869/j.issn.100-8551.2021.11.2441
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In order to research the small RNA variation of damage effect in rice caused by the γ-ray irradiation, radiation doses of 0, 300 and 400 Gy were applied to the rice seeds of Gaogengnuo, a small transcriptome was sequenced by using Ilumina HiSeqTM 2500 with three leaf stage seedling of irradiated GaoGengNuo. Result showed that total of 7 395 578 small RNAs, including 254 known miRNAs and 26 novel miRNAs were identified in 0 (CK) group; 10 315 701 small RNAs, 265 known miRNAs and containing 269 novel miRNAs were identified in the 300 Gy (Gy3) group; 6 469 869 small RNAs, containing 261 known miRNAs and 29 novel miRNAs were indentified in 400 Gy (Gy4) group. By Go analysis, the corresponding genes of differentially expressed miRNAs in Gy4 vs CK group were classified into 58 terms of three main GO categories, with the largest proportion of regions performing molecular processes; the differentially expressed miRNAs in Gy3 vs Gy4 group were classified into 7 terms of “biological process” and “molecular function” two main GO categories, among which ion binding, transition metal ion binding and oxidoreductase activity were the three most GO terms. Through analyzing the most significant enrichment 20 pathways with REGG pathway analysis, the corresponding genes of differentially expressed miRNAs in Gy3 vs CK group were majorly involved in plant pathogen interaction pathway, and the most significant pathway was plant circadian rhythm pathways; In Gy4 vs CK group, the highest number of genes were enriched in metabolic pathway, and the most significant pathway were glycerol phospholipids metabolism and ether lipid metabolism; In Gy3 vs Gy4 group, the highest number of genes were enriched in glycerol phospholipid metabolism, ribosome biogenesis in eukaryotes and endocytosis, and the most significant enrichment was ether lipid metabolism. By analyzing the variations of RNA level, it provides a new pathway to study the damage effect of rice seedling by irradiation.

Cloning and Expression Analysis of Transcription Factor PpWRKY18 in Peach
LI Sen, WANG Qingjie, CHEN Xiude, ZHANG Rui, LI Ling, DU Guiying, FU Xiling
2021, 35 (9): 1987-1993. DOI: 10.11869/j.issn.100-8551.2021.09.1987
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In order to explore the important role of WRKY transcription factors in plant resistance to adversity stress. This study obtained the PpWRKY18 gene in peach, and bioinformatics analysis showed that the PpWRKY18 protein contained a typical WRKY functional domain. Phylogenetic tree analysis showed that PpWRKY18 in peach was more closely related those in to Rosaceae species. PpWRKY18 was predicted to be a nucleolus localization protein, and was confirmed to be located in nucleus using transient expression in tobacco leaf through Agrobacterium tumefaciens injection. Fusion protein PGEX-PpWRKY18 was expressed after induction, and showed that the PpWRKY18 was mainly expressed in the inclusion body. Real-time PCR showed that the expression of PpWRKY18 was induced by drought, and the expression of PpWRKY18 decreased after rehydration. The protein phosphatase protein family (Protein Phosphatase 2C), C2H2 protein family (PpZAT5), ERF protein family (PpERF9), and BHLH protein family (PpBHLH92) were predicted to interact with PpWRKY18, therefore we speculate that PpWRKY18 regulates drought stress by interacting with these proteins, and may provide a theoretical basis for investigating the molecular mechanism of PpWRKY18 response to drought.

Traits Analysis of Offsprings Mutated by 60Co-γ Radiation in Iris hollandica
LIN Bing, FAN Ronghui, CHEN Yuhua, FANG Nengyan, YE Xiuxian, ZHONG Huaiqin, HUANG Minling
2021, 35 (10): 2205-2213. DOI: 10.11869/j.issn.100-8551.2021.10.2205
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In order to explore distinguishing main characters of mutant obtained by 60Co-γ radiation in Iris hollandica Zhanchi, botanical agronomic traits of 4 stable mutants (5-7CBS, 7-10SLZ, 7-10QLS, 10ZLL) and their wild-type Zhanchi were evaluated, the similarities and differences of the main ornamental traits (flowers) were analyzed and compared according to test guidelines of Bulbous Irises. It was found that mutants and wild type were basically consistent in plant morphology, flowering period and growth period. Among the 40 traits investigated, there were significant differences in the characteristics of five pseudo-qualitative characteristics between mutants and wild type, such as groud color of upper side of blade in outer tepal, color of inner tepal, color of upper side in bridge, color of upper side in crest, and groud color of upper side of claw in outer tepal. Another 35 character were highly similar in corresponding phenotype and values. The results showed that flower color of Iris hollandica has great genetic improvement potetiality with radiation breeding. It is speculated that flower color variation may be an important indicator for distinguishing mutants from wild type. According to the expression state of flower color, variation of flower color can be divided into four types: strengthening type, wild type, weakening type and full variant type. This study can provide basic data and reference for breeding new varieties in Iris hollandica.

Construction of Barley Mutant Population and Cytological Analysis of Leaf Width Mutation
QIN Dandan, XU Fuchao, XU Qing, PENG Yanchun, GE Shuangtao, DONG Jing, JIAO Chunhai
2021, 35 (2): 262-270. DOI: 10.11869/j.issn.100-8551.2021.02.0262
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To construct our own mutant library and provide materials for germplasm innovation and illustration of mechanisms involving in agronomic traits in barley and crops, mutant population of barley Edamai 934, which was an elegent cultivar bred by Hubei Academy of Agricultural Sciences, was constructed using 60Co-γ combined with EMS. Totally 982 mutants were identified, which showed significant and hereditable changes as compared with the wild type Edamai 934 on one of the following traits, leaf, stem, spike, seed, tiller, plant architecture, heading date, seed setting rate, and so on. Further cytological analysis of leaf width mutants showed that size and morphology of cells from flag leaf of different mutants varied a lot, suggesting that different mechanisms may involve in. This study lays foundation for barley functional genomics.
Research Progress in Traceability of Agricultural Products Using Stable Isotope
ZHAO Ruting, YANG Shuming, ZHAO Yan
2020, 34 (S1): 120-128. DOI: 10.11869/j.issn.100-8551.2020.34.0120
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With the enhancement of consumers' demands for the quality and safety of agricultural products, the traceability research of agricultural products keeps a good development trend. Stable isotope techniques are widely used in the research of agricultural products, including geographical origin traceability, authenticity identification and so on. In terms of geographical origin traceability, beef, mutton, milk, wine, honey, aquatic products, tea and grain have been investigated widely. It is difficult and important to narrow the scope of traceability area. The combination of stable isotope ratio with multi-element, amino acid and fatty acid content is the best way to narrow the traceability area. In terms of the authenticity of agricultural products, the variety and complexity of adulteration make it difficult for consumers to distinguish the true products. The fractionation of stable isotopes in agricultural products processing has been developed and gradually deepened. In this paper, the progress of the research on stable isotopes in the origin traceability, authenticity and fractionation process of agricultural products from 2015 to 2020 was reviewed. And the combination of stable isotope with other technologies for the geographical traceability of agricultural products was described. Furthermore, its deficiencies were summarized and prospected.
Determination of Main Volatile Substances in Strawberry Leaves and Its Inhibition Effects on Mycosphaerella fragariae (Tul) Lindau
TONG Zhichao, XU Yanqun, LI Dong, LI Li, LUO Zisheng
2021, 35 (2): 498-507. DOI: 10.11869/j.issn.100-8551.2021.02.0498
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In this study, Mycosphaerella fragariae was isolated and identified from strawberry leaves to explore the antibacterial effect of volatile substances in strawberry leaves on Mycosphaerella fragariae. The main volatile substances in strawberry leaves were determined and their effects on the spore germination, mycelial growth and mitochondrial membrane potential of the fungi were investigated. Results showed that C6 aldehydes and terpenoids were the two main volatile classes of strawberry leaves. Among them, hexanal, 2-hexenal,trans-2-hexenal and cis-3-hexenal, linalool and myrtenol were the dominant volatile compounds. Thus, these volatiles were selected to study the inhibition effect on the infection of M. fragariae of strawberry leaves by fumigation. On the concentration of 50 μL·L -1, linalool and myrtenol significantly inhibited the germination of spores and the growth of hyphae. Same effect was found for hexanal at 5 μL·L -1, trans-2-hexenal at 0.5 μL·L -1, cis-3-hexenal at 5 μL·L -1. Additionally, the mitochondrial membrane potential of the pathogen was significantly decreased, contributing to the growth inhibition on the fungi. Furthermore, the fumigation of the selected volatiles showed no damage to the strawberry leaves. These results indicated that with a suitable concentration the fumigation treatment of linalool, myrtenol, hexanal, trans-2-hexenal and cis-3-hexenal on strawberry leaves can effectively inhibit the infection of M. fragariae. This study provided a fundamental basis for the usage of plant volatiles as a new natural fungicide for plant disease control.
VIGS Silencing SlDCL2 and SlDCL4 Destroy Tomato Ty-1/ Ty-3 Resistance to Tomato Yellow Leaf Curl Virus (TYLCV)
YUE Ningbo, LI Yulong, SUN Yifan, PAN Pengcheng, PAN Yintao, ZHENG Yin, LI Yunzhou, LIANG Yan
2021, 35 (11): 2493-2500. DOI: 10.11869/j.issn.100-8551.2021.11.2493
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Tomato yellow leaf curl virus (TYLCV) seriously threatens the production of Solanaceae vegetable crops. Resistance markers Ty-1 and Ty-3 are a pair of allele genes, which are widely used in tomato breeding for TYLCV resistance. In order to explore the antiviral molecular mechanism of Ty-1/Ty-3, tomato ‘Y19’ with the Ty-1/Ty-3 resistance marker was used as the material, the key genes SlDCL2a/b/c/d (SlDCL2) and SlDCL4 involved in RNA interference (RNAi) were silenced by virus induced gene silencing (VIGS) technology, and their function in resistance of Ty-1/Ty-3 to TYLCV was study. The targeted fragments of SlDCL2 and SlDCL4 were amplified and sequenced, results showed that the silencing vector, pTRV2∶SlDCL2 and pTRV2∶SlDCL4 were constructed successfully. The silencing of tomato PDS (SlPDS) gene was used as a marker for successful silencing in plant, and the silencing efficiency of the SlDCL2 or SlDCL4 genes in the SlDCL2 or SlDCL4 silencing lines was detected by quantitative PCR (qPCR). The results showed that the expressions of corresponding gene in the SlDCL2 and SlDCL4 silencing lines were 50% less than that in the control plants, which indicated that SlDCL2 and SlDCL4 silencing vectors can indeed reduce the expression of corresponding genes. SlDCL2-silenced and SlDCL4-silenced plants were inoculated with TYLCV respectively. The results showed that the ‘Y9’ material had obvious symptoms of TYLCV, the disease severity of SlDCL2- and SlDCL4- silenced plants were 1.97±1.01, 2.35±0.75 respectively, significantly higher than that of the plants transformed with empty vector (0.14±0.24) and no vector (0.07±0.19). This result indicates that SlDCL2 and SlDCL4 play important roles in the resistance of Ty-1/Ty-3 to TYLCV, which laid a foundation for tomato breeding with resistance marker genes of Ty-1/3.

Construction of Ethyl Methane Sulfonate Mutant Library of Jinmai 47 and Preliminary Analysis of Quality Characteristics of High Generation Mutants
ZHANG Ting, WEN Hongwei, YUAN Kai, LU Lahu, SHI Xiaofang, ZHANG Mingyi, JI Hutai, YANG Bin
2021, 35 (8): 1731-1739. DOI: 10.11869/j.issn.100-8551.2021.08.1731
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To speed up creating of drought-resistant wheat resources and promote the genetic improvement of drought-resistant wheat varieties, the seeds of Jinmai 47 were treated with 0.6% EMS solution, a M 2 mutant library containing 12272 individual plants was constructed, and the mutation characters and mutation rates were investigated. In addition, the grain hardness index, protein content, wet gluten content, sedimentation value, water absorption, stability time, maximum resistance and extensive area of 50 M 7 and M 8 mutants selected in the previous study were analyzed. The results showed that: M 2 mutant library was rich in phenotypic variation, including stem character, fertility, spike type, disease resistance, leaf character, growth period and awn character, and the total mutation rate was 30.24%. Among which, the mutation rates of fertility, plant height and panicle shape were higher. There were not only morphological mutations in grain shape, grain length, plumpness and grain color, but also variations in grain quality characters of 50 mutants. Among them, the coefficient of variation of stability time, extensive area, maximum resistance and sedimentation value was the highest, the coefficient of variation of hardness index, protein content and wet gluten content was medium, and the coefficient of variation of water absorption was the lowest. Except for water absorption, positive variation occurred in a certain proportion of other characters. Four mutant materials with good quality characters were preliminarily screened, except hardness index and water γ-absorption, the other six quality characters were better than those of the wild type. This research provides valuable genetic materials for the discovery of wheat functional genes and new ideas for the drought resistance breeding program.
Mutagenic Effects of 60Co-γ Irradiation on Four Species of Lilium
LI Lihui, HU Yao, LEI Xingyu, ZHANG Yuelong, LI Honggao, HU Die, ZHANG Yong, DENG Gangqiao
2021, 35 (8): 1725-1730. DOI: 10.11869/j.issn.100-8551.2021.08.1725
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To investigate the mutagenic effects of different radiation doses of 60Co-γ rays on Lilium, the seedballs of Lilium lancifolium Thunb, Lilium bosniacum, Lilium henryi Baker and Lilium regale Wilson were used as test materials and treated with different dose(0,2,4,6Gy) of 60Co-γ rays. Statistical analyses of the germination rate, characters, physiological and biochemical indexes and variations.The results showed that the germination rate, plant height, bloomed rate and production of Lilium bosniacum, Lilium henryi Baker and Lilium regale Wilson gradually decreased with the increasing of radiation dosage. However, Lilium lancifolium radiated by 2 Gy showed no significant difference from the control,and by 4 Gy,the germination rate showed significant difference when compared with control. Under the radiation dose of 6 Gy,the seedballs could not sprout. 2 Gy dose radiation had little effect on the contents of chlorophyll for four varieties, 4 Gy treatment showed a downward trend.The activity of superoxide dismutase (SOD) increased first and then decreased in all varieties. While the malondialdehyde (MDA) increased gradually. The soluble sugar content of Lilium lancifolium Thunb increased first and then decreased, and other varieties increased with the increase of dose. The suitable radiation dose ranges for Lilium bosniacum, Lilium henryi Baker and Lilium regale Wilson were 2 to 4 Gy, 2 Gy and 2 Gy respectively. No appropriate dose was found for the Lilium lancifolium Thunb in this experiment. The results provide some technical references to a certain extent for Lilium radiation breeding.
Construction and Preliminary Screening of EMS Induced Mutant Library of Pumpkin ( Cucurbita moschata D.)
MIN Ziyang, HAN Xiaoxia, LI Yongqi, HU Xinjun, WANG Duanhua, SUN Xiaowu
2021, 35 (4): 761-768. DOI: 10.11869/j.issn.100-8551.2021.04.0761
Abstract163)   HTML1)    PDF (16267KB)(196)      
To accelerate the development of functional genomic research, the mutant library of Cucrbita moschata D. cultivar N87 was constructed. The seeds of the high-generation inbred line N87 were treated with ethyl methanesulfonate (EMS) solution of different concentrations for different times, and the phenotypic variation characteristics of pumpkin were identified.Considering the germination rate seedling rate and seedling growth,1.6% EMS for 12 h was selected as the appropriate treatment condition. Then 10 000 seeds of N87 were treated by 1.6% EMS for 12 h,and a pumpkin mutant library contained 725 M 2 families was constructed. The mutant library has rich variation in leaf, plant type, reproductive organ, fruit and other characters, with a total variation frequency of 11.2%. With constructing and analyzing the F 2 population from the cross of mutants and normal plants white flower mutation and short-internode mutation was found to be controlled by a single recessive gene respectively. The study first successfully created a pumpkin mutant population, which not only enriched pumpkin germplasm resources, but also provided useful mutant materials for pumpkin functional genomics research and breeding improvement.
Genetic Diversity Analysis of Iris Hybrid and Mutagenic Breeding Materials Based on SSR Markers
ZHOU Lin, CAI Youming, ZHANG Yongchun, YANG Liuyan
2021, 35 (3): 509-517. DOI: 10.11869/j.issn.100-8551.2021.03.0509
Abstract162)   HTML0)    PDF (1974KB)(238)      
In order to speed up the breeding process of Louisiana iris, hybridization and 60Co-γ radiation mutagenesis were carried out on five Louisiana iris varieties. Based on the EST-SSR markers of I.brevicaulis and I.fulva, 14 pairs of SSR markers were selected for genetic diversity analysis on Louisiana iris varieties, their hybrid offspring and mutagenic materials. The results showed that there was little genetic difference between the offspring of five Louisiana iris varieties and their maternal plants. However, after irradiation with 60Co-γ, great genetic differences from their parents were observed rays for five Louisiana iris cultivars. The survival rate of tissue culture plants was significantly negatively correlated with the dose of 60Co-γ mutagenesis. And the survival rates of different varieties under treatments of 2.5, 5, 10, 20 and 40 Gy were 20.77%~24.99%, 13.33%~17.89%, 10.11%~15.71%, 3.33~10.14% and 1.11%~2.77% respectively. The phenotypic traits of the mutagenic materials could be observed via tissue culture rapid propagation system. In this study, in vitro culture combined with 60Co-γ radiation mutation breeding was carried out for the first time on Louisiana iris, and 10 pairs of SSR markers were screened for identifying Louisiana iris varieties, hybrid offspring and mutagenic materials were screened. The research provide theoretical basis for Louisiana iris hybrid breeding, radiation mutation breeding and SSR-based germplasm identification.
Effects of Combined Application of Sheep Manure-Derived Organic Fertilizer and Chemical Fertilizer on Tobacco Growth and Soil Fertility
MA Yilin, WU Guanghai, SHEN Hongtao, WANG Xinzhong, ZHAO Shimin, MA Junhong, WANG Lihua, LIU Ling
2021, 35 (10): 2423-2430. DOI: 10.11869/j.issn.100-8551.2021.10.2423
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In order to investigate the influences of combined application of sheep manure-derived organic fertilizer and chemical fertilizer on tobacco (Nicotiana tabacum L.) growth and soil fertility, we used flue-cured tobacco variety Zhongyan101 as test materials and conducted a pot experiment to study the effects of different application ratios of sheep manure and chemical fertilizer(T0: 100% inorganic fertilizer N; T20: 20% sheep manure N+80% inorganic fertilizer N; T40: 40% sheep manure N+60% inorganic fertilizer N; T60: 60% sheep manure N+40% inorganic fertilizer N; T80: 80% sheep manure N+20% inorganic fertilizer N; T100: 100% sheep manure N) on tobacco agronomic characters, root morphological parameters, photosynthetic characteristics, soil nutrients and soil enzyme activities under equal amount of fertilizer application, and non-fertilizer application was set as a control (CK). The results showed that T0 and T20 treatments had higher soil available nutrients and could promote tobacco growth at 30 days after transplanting, but the contents of soil available nutrient and the growth potential of tobacco decreased significantly at 60 days after transplanting, and the content of soil organic matter and the activity of soil sucrase were also lower than that of other treatments at 60 days after transplanting. T80 and T100 treatments had higher soil organic matter content, but the activities of soil protease and urease, the contents of soil available nutrients and the growth of tobacco were lower than that of other treatments at 30 days after transplanting. T40 and T60 treatments had slightly lower soil available nutrients than that of T0 treatment at 30 days after transplanting, but the activities of soil sucrase, protease and urease, as well as the contents of soil available nutrient significantly increased at 60 days after transplanting, and could ensure tobacco growth at both 30 days and 60 days after transplanting. Compared with T0 treatment, T40 and T60 treatments increased the maximum leaf area by 11.9% and 10.7%, the shoot dry weight by 11.9% and 9.6%, the root dry weight by 12.1% and 26.8%, and the net photosynthetic rate by 17.6% and 16.2%, respectively, at 60 days of after transplanting. In general, the 40% and 60% sheep manure N application was more conductive to the growth of tobacco and the maintenance of soil fertility. This study provided a theoretical basis for the scientific application of sheep manure-derived organic fertilizer in tobacco-growing area of Luoyang.

Fine Mapping of Node Numbers on the Main Stem QTLs in Soybean Based on BAS and SLAF-Seq
YANG Yuhua, BAI Zhiyuan, WEI Baoguo, LEI Yang, ZHANG Ruijun
2021, 35 (9): 1953-1963. DOI: 10.11869/j.issn.100-8551.2021.09.1953
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To provide technical supports for marker assisted breeding and accelerate the cloning and functional verification of candidate genes of node numbers on the main stem in soybean, high throughput sequencing was used to detect the association region (QTL), fine mapping of the QTL were then conducted with the InDel markers developed based on the re-sequencing of parents. In this study, 102 recombinant inbred lines (RIL) derived from the cross between C025 with few node numbers on the main stem and Zhong119 with many node numbers on the main stem were used as experimental materials. Two mixed pools were constructed from 30 inbred lines with extremely few or extremely many node numbers on the main stem respectively. Five quantitative trait locus (QTL) related to the node number on the main stem of soybean were detected on chromosome 4 by the Bulked segregant analysis (BSA) and specific-locus amplified fragment sequencing (SLAF-Seq) high throughput sequencing method. In order to further narrow down the association regions, insertion-deletion (InDel) information between the association regions were obtained according to the re-sequencing of parental lines and the InDel markers were developed. The genotype of F 2 population was firstly analyzed by the InDel markers, and the major locus was mapped in the third associated region. Then eight co-dominant InDel markers were developed in the major region and all RIL lines were genotyped, nine individuals were obtained and the major region was divided into six exchange types. Combined with phenotypic analysis, the node number on the main stem was mapped between InDel markers Chr04-38 and Chr04-46, a region with only 171.9 kb, including six candidate genes. Therefore, the major locus of the node number on the main stem of soybean main stem was fine mapped. In short, the combination of BSA and high-throughput sequencing can effectively and rapidly detect the association regions of the node number on the main stem of soybean, furtherly combined the re-sequencing of parents to develop the tightly linked InDel markers can realize fine mapping of the major region. The developed InDel Markers Chr04-38 and Chr04-46 are tightly linked with the node numbers on the main stem of soybean, which can be used in the molecular marker assisted breeding for node numbers on the main stem in soybean.

Evaluation of the Introduction Performance of Ten Table Grape Cultivars in Weinan Area of Shaanxi Province
LI Yashan, LI Yumeng, REN Yizhao, LIU Xu, WANG Lujun, WANG Jinfeng, CUI Ping, WANG Yanjun
2021, 35 (12): 2756-2765. DOI: 10.11869/j.issn.100-8551.2021.12.2756
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In order to screen superior table grape cultivars that are suitable for cultivation in Weinan area of Shaanxi Province and optimize the planting structure of table grape cultivars, ten table grape cultivars including Xinyu, Flame Seedless, Zicui Seedless, Zaoheibao, Summer Black, Zitian Seedless, Hutai No.8, Jumeigui, Shine Muscat, Gold Finger were introduced for cultivation. Field observation and laboratory detection were used in the introduction experiment. The phenological phase and botany properties of the experimental cultivars were observed and recorded, physical and chemical compositions of berries were measured and sensory quality of berries was evaluated. Finally, the berry quality was evaluated comprehensively and ranked with the method of principal component analysis (PCA). The results showed that there were differences among the cultivars in the phenological phase, the botany properties, the physical and chemical compositions and the sensory scores under the conditions of simple rain shelter in Weinan area. The PCA analysis indicated that Xinyu and Hutai No.8 showed better quality, and the quality of Zaoheibao, Shine Muscat, Zitian Seedless were general, but the other cultivars were poor. Xinyu and Hutai No.8 were suggested to be the prior, and the cultivars of Zaoheibao, Shine Muscat and Zitian Seedless could be scaled up moderately. However, the other cultivars were not recommended to plant in large-scale in this area or the cultivation management for planting them had been improved. The results could provide some references for introduction of superior cultivars and planting structure optimization of table grape cultivars in Weinan area.

Identification and Analysis of Drought Related Genes in Foxtail Millet ( Setaria italica L.) at Germination Stage Based on Transcriptome Sequencing
DAI Xiaodong, ZHU Cancan, SONG Yinghui, WANG Chunyi, DAI Shutao, QIN Na, LI Junxia
2021, 35 (8): 1761-1770. DOI: 10.11869/j.issn.100-8551.2021.08.1761
Abstract154)   HTML12)    PDF (4805KB)(130)      
In order to identify the key genes responsive to drought stress and to elucidate the underlying mechanism in drought tolerance of foxtail millet, the germinating seeds of drought tolerant line Shanxi2010 and drought sensitive K359*M4-1 were subjected to RNA-seq before and post drought treatment. The data showed that 2300 and 3652 differentially expressed genes (DEG) were identified in Shanxi 2010 and K359*M4-1 respectively, including genes encoding zinc induced facilitator like protein, germination like protein, protein phosphorylase, transporter protein, late embryogenesis abundant protein (LEA), transcriptional factors, peroxidase and others. The GO (gene ontology) terms and KEGG (Kyoto Encyclopedia of genes and genomes) metabolic pathway enrichment analysis were carried out with the DEGs identified by different comparison groups. DEGs in these two lines were enriched in 52 and 21 biological processes, respectively. KEGG enrichment analysis indicated that DEGs were mainly enriched in the pathways including ‘starch and sucrose metabolism’, ‘plant hormone signal transduction’, ‘photosynthesis antenna protein’, ‘phenylpropanoid biosynthesis’, and ‘cutin, imine and wax biosynthesis’ and ‘secondary metabolite biosynthesis’. This study laid a foundation for excavating key genes related to drought tolerance and demonstrating the genes network that involved in drought responses in foxtail millet.
Effects of Three Protective Agents on Gardenia Irradiated by 60Co-γ Rays
LIU Liang, WANG Dan, WANG Cheng, CHEN Hong, HU Guoyu, LI Qing
2021, 35 (5): 1009-1019. DOI: 10.11869/j.issn.100-8551.2021.05.1009
Abstract153)   HTML2)    PDF (3565KB)(208)      
In order to study the effects of radiation mutagenesis and the protective effects of three protective agents on Gardenia jasminoides cuttings, the cuttings was firstly irradiated by 60Co-γ rays with different doses (5, 10, 20, 40 Gy), then soaked in three kinds of protective agents including sodium chloride (NaCl), cysteine (Cys) and vitamin C (Vc), with four different concentrations. After that, the indexes of rooting rate, survival rate, root length, number of roots, planting rate, internode length, number of branches, number of leaves, and leaf length, leaf width of gardenia cuttings had been analyzed. Damages of the cuttings gradually increased with the increase of radiation dose. The half-rooting dose of 60Co-γ rays for gardenia cuttings was 21.22 Gy; half-lethal dose was 81.65 Gy, and half-planting dose was 20.30 Gy. Among the treatments, 3.00 g·L -1 NaCl had good effects on rooting and growth of gardenia cuttings at 20 Gy, 0.12 g·L -1 Cys and 0.08 g·L -1 Vc had good effects at 40 Gy. All the three protective agents performed well on protecting the cuttings against damages under high dose 60Co-γ rays, the protective effects were dependent on radiation dose and the concentration of protective agents. This research can provide a certain technical reference for the radiation mutation breeding of gardenia and the application of protective agents.
Phenotypic Identification and Gene Mapping of Green Revertible White-Stripes Mutant wsl887 in Rice ( Oryza sativa L.)
YAN Xiaoqing, CHEN Nenggang, LI Huan, CHEN Feng, SONG Ze, YU Xianquan
2021, 35 (11): 2451-2462. DOI: 10.11869/j.issn.100-8551.2021.11.2451
Abstract152)   HTML16)    PDF (14770KB)(70)      

Discovery of thermosensitive rice leaf-color mutant genes can enrich rice genetic resources and providing materials for studying the mechanism of the chloroplast development regulated by low-temperature. In the present study, a new green revertible white-stripe mutant, wsl887, was isolated from Qiao-gang Pearl Rice, a local rice germplasm in Guizhou, by 60Co irradiation. Results showed that wsl887 displayed a white-stripe phenotype at seedling stage in the field. However, the main agronomic traits at maturation stage, didn't show significant differences from that of the wild type. When grown at 20℃ and 25℃, wsl887 displayed albino and white-stripe leaves, respectively, with significantly decreased photosynthetic pigment when compared with the wild type. Interestingly, when grown at 30℃, leaf color and photosynthetic pigment content of ws1887 showed no significant difference with those of the wild type. Transmission electron microscopy observation revealed that, wsl887 had no or deformed chloroplasts in mesophyll cells of the albino leaves at 20℃, but had the same number and normal morphology of chloroplasts at 30℃. The qRT-PCR results indicated that genes related to photosynthetic pigment metabolic pathway, photosynthesis and mitochondrial electron transport were differentially expressed in wsl887 at different temperature (20℃ and 30℃), when compared with those in the wild type. Genetic analysis demonstrated that the phenotype of wsl887 was controlled by a single recessive nuclear gene. Finally, WSL887 was located between SSR markers RM262 and RM5427 on chromosome 2, with a physical distance of 743.6 kb. By analyzing the genes located in the candidate region, wsl887 is a new thermosensitive leaf color mutant, which has great application value in hybrid rice due to its mutation without compromising the yield of the plant and the main agronomic traits under natural conditions. Under different temperature conditions, the expression of the genes in cytoplasm was significantly reduced in wsl887. Taken together, we hypothesized that the WSL887 could be the key gene responsible for the development of chloroplast in cytoplasm. This study laid a solid foundation for further cloning the wsl887 mutant gene and studying the gene function.

Polyploid Induction of Phalaenopsis Protocorms via Colchicine Treatment
WU Ting, ZHU Jun, YANG Jiahui, GE Hong, YANG Shuhua, ZHAO Xin, YU Xiaonan, JIA Ruidong
2021, 35 (11): 2463-2469. DOI: 10.11869/j.issn.100-8551.2021.11.2463
Abstract150)   HTML11)    PDF (5672KB)(48)      

In order to explore the effect of colchicine on chromosome polyploidization of Phalaenopsis, protocorms of Phalaenopsis H-03 (2n=2x=38) were used as the materials to induce polyploid by colchicine with mixed culture in this study. Colchicine concentrations in the medium were 0.05%, 0.1%, 0.2%, and the duration of co-culture was 5, 10, and 15 d respectively. The results showed that the most efficient condition for inducing mutation was treated with 0.05% colchicine for 15 d, which resulted in about 30.00% surviving plants and 50.00% mutated plants. The results measured by flow cytometry and root-tip squash indicated that a large number of chimeras among mutants after treatment. Compared with the control plant, the mutants showed dwarfed, with dark green, rough surface, broad leaves, short or round leaves. Some mutant plants also had abnormal and twisted leaves. The observation of stomata showed that the shape of stomata were rounder,the number of stomata per unit area decreased and the size of stomata are larger in mutants than that in the control, and the length and width of the stomata of mutants were increased by 76.45% and 38.99%, respectively. The results of this study provide a theoretical basis for breeding new varieties of Phalaenopsis.

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