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Cloning and Analysis of Two avrBs 3 /pthA Family Genes from JXOV Strain
XIA Geng-shou, ZHANG Yan
2014, 28 (12): 2167-2174. DOI: 10.11869/j.issn.100-8551.2014.12.2167
Abstract9394)      PDF (5155KB)(384)      

Two members of avrBs 3 /pthA family were cloned from Xanthomonas oryzae pv. oryzae(Xoo)race JXOV strain, and named as jva 1 and jva 2, respectively. We conducted the amino acid sequence analyses and phylogenetic analyses and constructed the prokaryotic expression vectors to investigate the gene function and toxicity of jva 1 and jva 2. The results showed that the deduced amino acid sequences of Jva1 and Jva2 shared typical characteristics with other gene members of AvrBs3/pthA family. The molecular mass of proteins jva 1 and jva 2 from BL21(DE3) is about 100 KD on SDS-PAGE which is similar to presumed monomer protein. Dimerization of jva 1 or jva 2 did not occur in prokaryotic cell BL21, although the dimerization of AvrBs 3 was independent of plant proteins; Neither jva 1 nor jva 2 enhanced the PXO99Astrain virulence on fifteen rice cultivars holding different resistance genes, The gene functions of jva 1 and jva 2 need further investigation. These results provided important basis for understanding mechanism of pathogenicity of Xoo on rice and gene functionary diversity of members of related gene family.

Expression Pattern and Stress Responses of Wax Biosynthesis Related Gene OsGL1-5 in rice
ZHOU Ling-yan, NI Er-dong, ZHU Li-ya, LIANG Hong, ZHUANG Chu-xiong
2013, 27 (7): 938-945. DOI: 10.11869/hnxb.2013.07.0938
Abstract7622)      PDF (1097KB)(532)      

In this study, the promoter of wax biosynthesis related gene OsGL1-5 in rice was analyzed via bioinformatics, and the temporal and spatial expression of OsGL1-5 was analyzed by semi-quantitative RT-PCR and mRNA in situ hybridization using the different tissues and developmental stage panicles as materials. In addition, the expression of OsGL1-5 was analyzed by semi-quantitative RT-PCR when the seedlings were treated with stresses, including 200mmol·L-1 NaCl, 100μmol·L-1 ABA and 1.0% H2O2. The bioinformatics analysis showed that the promoter of OsGL1-5 (about 2 kb upstream of OsGL1-5) was predicted to contain important regulatory elements including drought, high salt, coldness, ABA, light regulation, copper, and oxygen responsive elements. The analysis of OsGL1-5 expression profiles revealed that OsGL1-5 was expressed highly in panicles at different developmental stages and specifically expressed in the trichomes of glume and the tapetum of anther, low expression of OsGL1-5 was also detected in the leaves, however, its expression was not detected in the stems and roots. The analysis of OsGL1-5 in response to stresses showed that the expression of OsGL1-5 was increased at different time when the seedlings were treated with NaCl, ABA and H2O2.

ZHANG Jia-qi1 ,2 ZHU Jin-qi2 LI Sheng2 WEI Zhao-jun1
2010, 24 (5): 926-931. DOI: 10.11869/hnxb.2010.05.0926
Abstract6012)      PDF (2068KB)(1153)      

Trypsin-modulating oostatic factor (TMOF) can inhibit trypsin activity in insect midgut dramatically. To analyze inhibitory ability of Aedes aegypti TMOF analogues genes to insect, we reconstructed plant expression vectors containing Aedes aegypti TMOF analogue genes, and then transformed those into tobacco. The synthetic Aea-TMOF analogues were cloned into modified PCAMBIA1301 vector that contains constitutive promoter CaMV35S. The recombinant plasmids of PCAMBIA1301+Aea-TMOF were transferred into Agrobacterium tunefacien LBA4404 by freeze-thaw method; and then were used to infect tobacco leaf discs. Using PCR and GUS methods, we proved that the Aea-TMOF analogue genes were integrated into tobacco genome.

Yang Changju Liu Ganming Deng Wangxi Yang Zhihui Hu Jianfang(Department of Plant Protection,Central China Agricultural University,Wuhan 430070)
1993, 7 (02): 88-92. DOI: 10.11869/hnxb.1993.02.0088
Abstract5611)      PDF (284KB)(987)      

4-5 days old male pupae of Indian meal moth,Plodia interpunctella Hubner were irradiated with 1,3,5,7 and 10 krad of 60Co γ-ray The male moths developed from treated pupae and their F1 generation were put together with normal female moths for copulation separately.The genetic effects of irradiation was studied and the results showed that the effects of irradiation were significant,which related to the irradiation dosage,on both parental and filial generations.Only 40% of the pupae was emerged,when they were irradiated at 10 krad.The effects on deformation rates,survival rates,copulation abilities,fecundities,hatching rates and genetic sterilities varied with different irradiation dosage treatment.With consideration of the total irradiation effect,dosage lower than 5 krad is desirable for inducing the sterility of adults and sex chain recessive lethal gene.With 1,5 and 7 krad of irradiation,a mutant of P.interpunctella with transparent wings was induced,which provides a marking feature in control of Indian meal moth by sex chain recessive lethal gene.

ZHAO Feng wu 1 LI Hui min 1 MA Jun yong 1 ZHAO Ming 2 ZHANG Gui fang 2 WANG Mei yun 2 (1.Dry Land Farming Institute, HAAFS, Hengshui, Hebei, 053000; 2.China Agricultural University, Beijing, 100094)
2004, 18 (05): 335-339. DOI: 10.11869/hnxb.2004.05.0335
Abstract5453)      PDF (315KB)(1304)      

A three generic hybridization was made from upland rice genotype Yuan F2 1 derived from a wild hybridization of upland rice Han 65 (Oryza sativa) /Long awn barnyard grass (Echinochloa caudata) , with a sorghum genotype of Shennong 133 (Sorghum bicolor) . Successive studies of its F\-0, F\-1 and F\-2 showed that: (1)F\-0 was highly cross\|incompatible and accompanied with seed abortion with seed set rate of 2\^63%; (2)F\-1 had excellent fertility with good heterosis. The characteristics such as plant height, head length, first branches, florets, seeds/head, seed set rate, 1000\|grain weight and leaf area of main stem in F\-1 were 0\^61%~20\^56% over its female; and leaf number was 1 piece more than its female. Additionally, it also showed red awn, red glume and purple stigma, which was different from its female's white color; (3) characters segregating did happen in F\-2, besides agronomic traits such as plant height, ear length, etc, the typical segregating were from head type, awn length, awn color, stigma color and glume color.

WANG Cai bin CHENG Bo SUN Xiu shan ZHENG Ya ping CHEN Dian xu (Shandong Peanut Research Institute, Qingdao Shangdong prov. 266100)
2002, 16 (02): 98-102. DOI: 10.11869/hnxb.2002.02.0098
Abstract4990)      PDF (260KB)(1122)      

Under the condition of wheat peanut(summer) cropping system, the nitrogenous fertilizer utilization ratio(NFUR) was 32 52% when the total annual nitrogenous fertilizer was applied to both crops by three times (base and top dressing to wheat and base fertilizer to peanut, M 3) and was 37 01% when the total annual nitrogenous fertilizer was applied to wheat by twice(base and top dressing to wheat, M 2). The nitrogen fertilizer recovery ratio was 69 24% in M 3, 12 03% higher than that in M 2. The yields of wheat and peanut in M 3 were 7 7% and 3 4%, respectively, higher than that in M 2, showing that the M 3 was ideal fertilizer application mode under the wheat peanut cropping system. The NFUR was 16 08%~35 63% for wheat and was 23 70% for peanut. The NFUR was 6 57%~7 70% for wheat with the base fertilizer and was 10 03%~12 73% for wheat with top dressing fertilizer (elongation stage). Great effects of fertilizer applying time and rate on the NFUR were observed.

Fine Mapping of QTL qtgw1 for Thousand-Grain Weight of Rice
LIN Wen-chun, YU Shou-wu, RUAN Guan-hai, FAN Ye-yang, XIE Jian-kun
2014, 28 (2): 217-223. DOI: 10.11869/j.issn.100-8551.2014.02.0217
Abstract4941)      PDF (1139KB)(328)      
Grain weight is one of the major components of rice yield-associated traits, which is also an important factor effecting grain quality trait. The aim of this study is to finely map a quantitative trait locus (QTL) for thousand-grain weight (TGW). Five residual heterozygous lines (RHLs) carrying heterozygous segments of RM151-RM10404, RM151-RM10425 and RM1344-RM5359 in the almost pure parental background were selected from the recombinant inbred lines of Zhenshan 97/ Miyang 46. F 2:3 populations of the five RHLs were grown for QTL analysis for TGW,the target region RM151-RM1344 were confirmed existing a QTL for TGW of rice, and the allele for increasing TGW is derived from the parent Zhenshan 97. Using simple sequences repeat marker (SSR) analysis, four individual plants with the heterozygous segments of RM151-RM3746, RM151-RM10402, RM10381-RM10425 and RM10381-RM10425 were selected to develop F 2 populations (FF population), and mapped the qTGW1 between RM10376-RM10404. Three individual plants with the heterozygous segments of RM10381-RM10402,RM10390-RM10425 and RM1344-RM10425 were used to develop their F 2 populations (L population). Using these four F 2 populations for further fine mapping, we narrowed the qtgw1 between RM10381 and RM10404 with a physical distance of about 246.6kb.
Orange Storage Time Determination Based on Electronic Nose Combined with Surface Acoustic Wave Resonator
WANG Min-min, ZHAO Meng-tian, DU Jia-su, HUANG Jie, CHENG Si-yi, YIN Fang-yuan, HUI Guo-hua
2014, 28 (4): 647-654. DOI: 10.11869/j.issn.100-8551.2014.04.0647
Abstract4844)      PDF (1264KB)(254)      

In this study, orange storage time determination is conducted using electronic nose (EN) combined with 433MHz surface acoustic wave resonator (SAWR). Responses of EN and SAWR to orange samples under different storage time are measured. Weight loss percentage of orange samples is also measured. Orange storage time predicting models based on EN, SAWR, and EN combined with SAWR are developed. Experimental results demonstrate that weight loss percentage index increases with the increase of storage time. Predicting model based on EN combined with SAWR presents higher orange storage time predicting accuracy than the other two models. Orange storage time predicting system using EN combined with SAWR has unique advantages including fast response, high accuracy, low cost, etc. This method provides a novel way for fruit quality analysis.

LI Li-qin
2011, 25 (3): 461-468. DOI: 10.11869/hnxb.2011.03.0461
Abstract3407)      PDF (1361KB)(1728)      

In higher plants, WRKY gene family plays significant role in transcriptional regulation. They are widely involved in biotic stress, abiotic stress, growth, development and metabolism regulation. In this study WRKY12 from tobacco was cloned by homology cloning. The deduced protein sequence showed that this protein belonged to the second group of WRKY family and the zinc-finger structure is C-X4-C-X23-H-X1-H. Phylogeny results showed WRKY12 was much closer to GmWRKY6 and GmWRKY12 from bean, generated 50% and 45% amino acids similarity. Northern Blot experiment showed that the gene expression was induced at 4℃ treatment. By SDS-PAGE analysis, the best expression condition of WRKY12 was obtained when the host bacteria- E.coli Rosetta-gami B (DE3) was grown for 8h at temperature 18℃ added 0.2mmol/L IPTG. Further experiment generated the purified protein and antibody with high specificity.

Influence of Water Management and Tillage Patterns on Nitrogen Utilization of Rice
XU Shi-Hong, LIANG Tian-Feng, ZENG Hua-Zhong, JIANG Li-Geng, DING Cheng-Quan, ZHANG Yu
2009, 23 (6): 1065-1069. DOI: 10.11869/hnxb.2009.06.1065
Abstract3175)      PDF (1959KB)(1386)      

Taking the rice vatieties of Jinyou 253 and Guihan 1 as the material, pot experiments were conducted to evaluate the differences of nitrogen utilization by rice using 15N tracer techniques in the early and late seasons of 2008. Experiment was carried out under three water management modes (Wetting irrigation, alternation irrigation and water layer irrigation) and two soil tillages patterns (no tillage and conventional tillage). The results showed that: (1) Wetting irrigation reduced the nitrogen uptake from fertilizer by rice plants under both soil tillage patterns, and the differences were significant for the basal and tillering fertilization. (2) More nitrogen from fertilizer was absorbed by rice plants and less was reserved in soil under wetting irrigation condition. (3) Nitrogen uptake by rice plants under wetting irrigation condition was obvious lower than that of rice with alternation or water layer irrigation under the conventional tillage pattern, but the influence of wetting irrigation on nitrogen uptake was less under no tillage pattern. (4) Grain yield and nitrogen grain production efficiency were reduced under Wetting irrigation condition.

Irradiation Sensitivity of 6 Cultivars in Euphorbia pulcherrima
SU Jia-Le, LI Chang, JIANG Wei-Hua, LIU Xiao-Qing, CHEN Lu
2009, 23 (6): 990-993. DOI: 10.11869/hnxb.2009.06.0990
Abstract3163)      PDF (2520KB)(1451)      

The cuttings seedlings of 6 Euphorbia pulcherrima cultivars were irradiated by 60Coγ-rays, and the effects on the survival rate and plant growth were investigated. Then the regression models were used to analyze relationship between the characters change and doses. The results showed that the growth of Euphorbia pulcherrima were decreased after irradiation and the effect became stronger with the increase of irradiation dose. The indexes of survival rate, plant height, leaf length, leaf width and leaf area were all showed significant negative correlation with the irradiation doses. The medial lethal doses (LD50) of 6 Euphorbia pulcherrima cultivars were from 21.94 Gy to 28.71 Gy, and the radiosensitivity was different among tcultivas.

Mapping of Fertility Restoring Gene for Male Sterility With Ae.kotschyi, Ae.variabilis, Ae.ventricosa Cytoplasms by Using SSR Markers in Wheat
GUO Yan-Ping, ZHANG Gai-Sheng, CHENG Hai-Gang, SHU Zhan-Wang, ZHANG Long-Yu, NIU Na, MA Shou-Cai, LI Gong-Xia
2009, 23 (5): 729-736. DOI: 10.11869/hnxb.2009.05.0729
Abstract3112)      PDF (433KB)(1617)      

Microsatellite markers were employed to map the major restoring gene for male sterility with Ae.kotschyi, Ae.variabilis, Ae.ventricosa cytoplasms in wheat by using materials of NILs and BC1' to probe into the genetic mechanism of fertility restoration. The results showed that 18 sets of SSR primers which were located on the chromosome 1BS of wheat were screened for polymorphism between the NILs, and 6 of which were found polymorphic. Linkage analysis indicated that Xwmc406,Xbarc8,Xwmc611 and Xgwm273 were linked to the restoring gene. The breeding for new fertility restorer lines of male sterility with Ae.kotschyi, Ae.variabilis, Ae.ventricosa cytoplasms in wheat would be facilitated by using these polymorphic markers. The result also showed that the fertility restoration of male sterility with Ae.kotschyi, Ae.variabilis, Ae.ventricosa cytoplasms in wheat was mainly controlled by both major fertility restoring gene and some minor enhance genes of fertility. The restoring gene which was marked in this research should be Rfv1. The distances between the restoring gene and the 4 markers are 2.7, 2.8, 21.4 and 26.2cM, and the sequence of the 4 markers and Rfv1 were Xwmc406, Rfv1, Xbarc8, Xwmc611 and Xgwm273.

Development and Application of Maize Inbred Line Luyuan 92
ZHOU Zhu-Hua, XU Li-Hua, WANG Li-Li, XU Fang-Zuo, XING Yan-Ju, ZHANG Feng-Yun, QIU Deng-Lin, YIN Wei-Jun, HAN Jin-Long, XU Xiang-Bo, DING Yi
2009, 23 (6): 986-989. DOI: 10.11869/hnxb.2009.06.0986
Abstract3049)      PDF (1644KB)(1384)      

The inbred line Luyuan 92, which was bred by irradiation technology and conventional method, is with many good traits, such as high combining ability, strong stress resistance, high yield and good agronomical charactors. It is one of the key inbred lines in China. 19 varieties bred using Luyuan 92 directly or indirectly had been passed national or provincial approval. According to the incomplete statistics of seed management departments in some provinces, the varieties from Luyuan 92 had been planted 5.2×106 hm2, the economic benefit reached RMB 18.82×108 till 2008.

ZHANG Jing-Xin, YANG Qi-Yun, WANG Hui, CENG Li-Xian, LIU Yong-Zhu, GUO Chao, SHU Xiao-Yuan, CHEN Zhi-Jiang
2010, 24 (3): 425-429. DOI: 10.11869/hnxb.2010.03.0425
Abstract3044)      PDF (132KB)(1482)      

To screen the resistance lines to rice blast, the blast resistance of SP3 and SP4 progenies derived from rice variety Hanghui 7 were evaluated after satellite flight, and the genomic DNA polymorphism of the resistant variants selected from SP3 was compared with the wild type by microsatellite markers. The results indicated that the SP3 Variant line H24, which was selected from the 250 space-induced lines (SP3 ) with excellent agronomic and economical characters, showed resistance segregation (119R∶108S) against blast isolate GD3286. It was demonstrated that the resistance of H24 might be controlled by two dominant and complementary resistance genes. The resistance of H24 was still segregated in SP4, but the resistance spectrum of H24 was 84.4% in SP5,much higher than the wild type, 40.6%, and H24 especially showed resistant against some blast isolates of broad pathogenic spectrum or specialized pathogenicity;further more, the DNA polymorphism wasn’t detected between H24 and its wild type by 229 SSR(simple sequence repeat) markers covering the rice genome equally.

Screening and Identification of HMW-GS Mutants in Wheat
LI Peng, SUN Ming-Zhu, ZHANG Feng, ZHANG Feng-Yun, LI Xin-Hua
2009, 23 (6): 935-938. DOI: 10.11869/hnxb.2009.06.0935
Abstract2992)      PDF (1415KB)(1488)      

The variation and compositions of high molecular weight glutenin subunits (HMW-GS) in the SP2 of space loaded Baiyingdong 2 were analyzed by SDS-PAGE techniques. 2 mutants (mut1 and mut2) of HMW-GS were screened out, the variation frequency was 2.17%. Using gliadin protein electrophoresis proved the HMW-GS mutants were induced by space-flight mutation. 4 kinds of HMW-GS compositions were found in mut1 seeds, and the mutations occurred in Glu-1B and Glu-1D sites. The mutated subunits resulted from the coding genes’ mutations which were induced by space-flight, their electrophoretic mobility changed significantly, these mutated subunits were new glutenin subunits. The results of this study showed that space-flight mutation could create new HMW-GS, and space-flight mutation was an efficient method for enriching the HMW-GS gene resource.

WANG Ying-jun, DENG Shi-huai, JIANG Jing, SHANG He, LIN Bo, SUN Ya-qin, HU Xiao-mei
2011, 25 (3): 588-593,581. DOI: 10.11869/hnxb.2011.03.0588
Abstract2989)      PDF (435KB)(1745)      

A pot experiment was conducted to study the effects of simulated acid rain on foliar damage, chlorophyll (Chl) content, membrane permeability (MP), antioxidant enzyme activity and gas exchange parameters in leaves of 2-year H. mitabilis seedlings. Four gradients of simulated acid rain solutions of pH 5.6 (CK), 4.0, 3.0 and 2.0 were used. Results showed that the leaves treated by pH 2.0 and 3.0 showed visible injury spots to various extent. The membrane permeability (MP) increased significantly after acid rain stress with an extremely significant negative correlation to pH value (r=-0.961**). The maximum catalase (CAT) activity and peroxidase (POD) activity were observed at pH 2.0(48.75mg·min-1·g-1·FW and 77.69U·min-1·g-1·FW, respectively), while that of superoxide dismutase (SOD) activity were observed at the pH 3.0(216.80U·g-1·FW). Chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll content, Chl a/b, net photosynthetic rate (Pn), stomatal conductivity (Gs) and transpiration rate (Tr) decreased with decreaseing pH value of pH, and intercellular CO2concentration (Ci) showed a trend of decreasing first and then increasing, while water use efficiency (WUE) was opposite. The visible damage threshold of H. mitabilis by simulated acid rain was less than pH 2.0, and the threshold on the physiological activity was among pH 2.0 to 3.0, which indicated that H. mitabilis had higher tolerance to acid rain stress. This study could provide theoretical reference for cultivation management and resistance screening of H. mitabilis, and offer basic theory to choose the greening tree species in Southern cities.

Breeding of Rice Transgenic Sterile Lines with Bore Resistance by Small Population Backcrosses Combined to GUS Marker-assisted Selection
ZHOU Yi-Feng, ZHAO Fei, CUI Hai-Rui, SHU Qing-Yao, SHEN Sheng-Quan
2009, 23 (6): 905-910. DOI: 10.11869/hnxb.2009.06.0905
Abstract2977)      PDF (180KB)(1877)      

With D62B taken as recurrent line and Tb26 derived from“KMD”as transgenic insect-resistant donor line, we improved the borers resistant characteristics of D62A(B).Bt-type D62A(B) were constructed by consecutive backcrosses combined to GUS molecular marker-assisted selection. F1 hybrids derived from Bt-D62A also had high borer resistance.Bt-type D62A had the characteristics of good flower organ and flowering behavior, complete pollen sterility.Otherwise,gronomic performance,quality traits and combining ability were the same as that of D62A(CK).If D62A could be gradually replaced for Bt-type D62A, we could save the cost of pesticide, reduce pollution and so on.

The Genetic Transformation of Strawberry with Winter Flounder Antifreeze Protein Gene
SUN Rui-Fen, LI Tian-Ran, AN Yu-Lin, LI Kun, SHI Hui-Qin, NIU Su-Qing, ZHAO Jing-Hui
2009, 23 (3): 423-428. DOI: 10.11869/hnxb.2009.03.0423
Abstract2976)      PDF (358KB)(1247)      

Antifreeze protein (AFP) of Pseudopleuronectes americanus gene encoding pre,pro and mature was introduced into strawberry (Fragaria X ananassa cv. Xingdu 2 and Allstar) by mediated Agrobacterium gene transfer of a binary vector, and the axenic leaf discs and lower part of petioles of strawberry were used as explants.The results showed that the foreign AFP gene encoding pre,pro and mature was integrated independently into the genome of six transgenic strawberry plants by PCR and PCR-Southern bloting analysis of seven Kmamycin-resistant transformed plants.Three regenerated Xingdu 2 lines were positive for pre,pro and mature,and their transformation efficiency was 1.56%,1.49% and 1.67% respectively.Three regenerated Allstar lines were positive for mature and its transformation efficiency was 4.05%.

ZHANG Zhong-Gui, LIU Bin-Mei, HU Hua, ZHANG Li-Li, WANG Min, TUN Ti-Jin
2010, 24 (3): 430-435. DOI: 10.11869/hnxb.2010.03.0430
Abstract2947)      PDF (950KB)(2708)      

Fourteen plant height mutants of Nipponbare were used to study the effect of plant height on the agronomic characters and lodging resistance. The results indicated that the plant height was positively correlated with spike length, third internode length,height of gravity center, fresh weight of main stem, dry weight of main stem, thousand-grain weight, grain-yield per plant and biological yield, and the second internode length.Meanwhile, plant height played an important role in lodging resistance, it was significantly positively correlated with lodging index and negatively correlated with bending moment and culm type index.The correlation between agronomic characters and lodging resistance showed that several agronomic characters had strong impact on the lodging resistance,such as spike length, height  of gravity center, basal internode length(first and second internode), fresh and dry weight of main stem, dry weight of basal internode, seed setting, thousand-grain weight, grain-weight per plant and biological yield.

Characteristic,Inheritance and Breeding Application of Rice Mutants with Greenable Albino Leaf
FANG Xian-Tao, MA Hong-Li, ZHAO Fu-Yuan, ZHANG Qing-Qi, ZHANG Shu-Biao
2009, 23 (1): 1-6. DOI: 10.11869/hnxb.2009.01.0001
Abstract2924)      PDF (226KB)(1805)      

Inheritance and main agronomic traits of photo-thermo-sensitive genic male sterile line with green-revertible albino leaf were investigated. The results indicated that the mutants may be divided into three types:albino regreening type(W2, W3, W4 and W10) , albino to kelly type(W9) and abino-regreening-albino-regreening type(W1 and W7). Genetic study indicated that the green-revertible albino leaf color trait of the mutants were controlled by a single recessive gene.These mutants had similar agronomic traits and fertility characteristics to the corresponding male sterile line Peiai 64S. The hybrids of these mutants had similar characteristics with original-hybrids in plant type, developing of tillers and plant height. The yield components of these mutants hybrids were different depended on different mutants. The yield potential of hybrids of W1,W2 and W3 were similar to the original-hybrid. The results also indicated that W1, W2 and W3 have breeding application value.

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