Table of Content

12 February 2016, Volume 42 Issue 02

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

An Integrated Genetic Linkage Map from Three F₂ Populations of Cultivated Peanut (ArachishypogaeaL.)

GUO Jian-Bin,HUANG Li,CHENG Liang-Qiang1,CHEN Wei-Gang,REN Xiao-Ping,CHEN Yu-Ning,ZHOU Xiao-Jing,SHEN Jin-Xiong,JIANGHui-Fang

Acta Agron Sin. 2016, 42(02):  159-169.  doi:10.3724/SP.J.1006.2016.00159

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The genetic linkage map is important for peanut molecular breeding. Construction of integrating genetic linkage map using multiple populations is an effective approach to increase the marker density of map. Three maps were constructed with three F₂ populations, respectively in the present study. Based on anchored SSR markers in the three maps, we constructed anew map with 792 SSR loci and total map distance of 2079.5cM. The length of linkage groups varied from59.1 to175.8cM, and the number ofmarkers wasfrom20 to 66 inthe integrated linkages groups.Comparingthe intervals of QTLslinked to the pod size and seed size in the three F₂ populations withthe markers in the integrated linkage groups, all the QTLs linked to the pod size and seed size could be found in the integrated map. Some intervals of QTLs had more markers in the integrated map than in the F₂ linkage groups in the present study. The markers in the intervals of QTLs of the integrated map could be used for fine mapping.

Molecular Cloning of ZmPP6C Gene and Its Expression Patterns in Response to Light and Stress Treatments in Maize (Zea mays L.)

YUAN Huan-Huan,SUN Guang-Hua,YAN Lei,GUO Lin,FAN Xiao-Cong,XIAO Yang,MENG Fan-Hua,SONG Mei-Fang,ZHAN Ke-Hui,YANG Qing-hua, YANG Jian-Ping

Acta Agron Sin. 2016, 42(02):  170-179.  doi:10.3724/SP.J.1006.2016.00170

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PP6C is the catalytic subunits of Ser/Thr protein phosphatase 6 (PP6) gene, which plays important roles in auxin transport polarity, ABA (abscisic acid) signal transduction, flowering time control though light signaling pathway. To clarify structural characteristics of PP6C protein and the evolution relationships among plant PP6Chomologs, we cloned ZmPP6C gene by RT-PCR. The open reading frame (ORF) of ZmPP6C possesses 912 nucleotides and encodes 303 amino acid residues with one PP2Ac domain (the catalytic subunits of Ser/Thr protein phosphatase 2A). Phylogenetic analysis indicated that ZmPP6C belongs to the same branch with the PP6C of Sorghum bicolor, and shows high similarity to all PP6C proteins from other monocotyledons and dicotyledons. Further quantitative RT-PCR (qRT-PCR) assays indicated that ZmPP6C was highly expressed in leaf and lowly in stem, stamen, pulvinus, sheath, and pedical. ZmPP6C transcription abundances could respond to different light and circadian treatments (both long-day and short-day conditions), especially to the light transitions from the dark to far-red or red light condition. In addition, ZmPP6C transcription abundances were up-regulated by high osmosis, high salt and water logging. Our results suggested that ZmPP6C may be involved in light signaling pathway, flowering time control, and abiotic stress response in maize, and its roles in crop improvement are worthy of more exploration in the future.

Genetic Diversity and Association Analysis of Agronomic Characteristics with SSR Markers in Hulless Barley

MENG Ya-Xiong,MENG Yi-Lin,WANG Jun-Cheng,SI Er-Jing,ZHANG Hai-Juan,REN Pan-Rong,MA Xiao-Le,LI Bao-Chun,YANG Ke,WANG Hua-Jun

Acta Agron Sin. 2016, 42(02):  180-189.  doi:10.3724/SP.J.1006.2016.00180

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The objectives of this study were to find molecular markers associated with yield-related traits and guide parental combination in molecular marker-assisted breeding and hybrid breeding of hulless barley (Hordeum vulgare L. var. nudum HK. f.). A natural hulless barley population composed of 108 parental varieties/lines was screened with 92 SSR markers, in which 48 markers were polymorphic. Population structure was analyzed based on the polymorphic SSR data and association between markers and five agronomic traits were performed in TASSEL GLM (general linear model) and MLM (mixed linear model) programs. A total of 156 alleles were detected in the 108 varieties/lines with 2–6 alleles per locus. The Shannon’s index of the population ranged from 0.6727 to 1.1368 and the genetic similarity between varieties ranged from 0.2250 to 1.0000, with the mean of 0.7585. Structure analysis revealed four genetic subpopulations for the entire materials tested. Based on GLM analysis, 12 SSR markers were found to be associated with plant height, spike length, grain number per spike and tiller number, with phenotypic contributions of 11.5%–17.6%, 19.4%–45.4%, 15.4%–22.1% and 29.2%, respectively. Based on MLM analysis, 8 SSR markers were associated with plant height, awn length, and spikelet compactness, with the phenotypic contributions of 31.71%–49.88%, 28.1%–37.2%, and 22.7%–32.7%, respectively. These associated markers were distributed on 6 chromosomes of the barley genome.

Cloning and Functional Analysis of Polygalacturonase Genes from Ciboria shiraiana

LI Meng-Jiao,Lü Rui-Hua,YU Jian,CAI Yu-Xiang,WANG Chuan-Hong,ZHAO Ai-Chun,LU Cheng,YU Mao-De*

Acta Agron Sin. 2016, 42(02):  199-200.  doi:10.3724/SP.J.1006.2016.00190

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Polygalactuionase (PG) is a kind of cell wall structural proteins that can catalyes the decompocition of alpha- (1,4)-polymer of galacturonia acid , which makes the cell wall structure disintegration and fruit softening. In the paper, we analyzed the expression patterns of PG genes in the leaf infection by the hypha of C. shiraiana at different growth stages by using qRT-PCR methods. The results showed that the cDNA of CsPG1 gene was amplified from the sclerotia of C. shiraiana by RT-PCR, namely CsPG1 (GenBank accession number: KR296662) with 1143 bp of full length, encoding 380 amino acid residues. On the basis of the highest level of gene expression in the process of infecting rape leaf, the main gene of the CsPG gene family was confirmed. CsPG1 was cloned into pET-28a(+) vector and expressed in E. coli BL21 (DE3). The recombinant CsPG1 protein was expressed in the form of inclusion bodies without activity towards polygalaturonic acid. The optimal urea concentration for dissolving CsPG1 inclusion bodies was 6 mol L^–1, CsPG1was renaturated by dilution gradiently at low temperature. We sorted out single band after purified by High-Affinity Ni-NTA Resin, and obtained soluble protein. The specific activity of renatured CsPG1 was 5.02 U mg^–1. The result of biological test showed that the recombinant protein accelerated the fruit maturity and softening of Jialing 40 (Morus atropurpurea Roxb.). So we speculated that the CsPG1 protein is related to the infection of C. shiraiana to mulberry fruits. The result reveals the sensitivity difference among mulberry varieties to sclerotial disease, providing the molecular evidence for preventing and controlling the sclerotial disease in mulberry cultivation.

Genome-wide Identification and Function Analysis of SBP Gene Family in Maize

PENG Hua,HE Xiu-Jing,GAO Jian,LUO Mao,PAN Guang-Tang,ZHANG Zhi-Ming

Acta Agron Sin. 2016, 42(02):  201-211.  doi:10.3724/SP.J.1006.2016.00201

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SBP gene family, as a plant special transcription factors is involved in plant growth and development, as well as many physiological and biochemical processes. Recently, SBP transcription factor family has been identified in model plants, such as Arabidopsis and Oryza sativa; however, systematic analysis of SBP transcription factor family in maize (Zea mays L.) is scarcely. In this study, based on homology alignment technology, we aligned all known SBP TFs from Arabidopsis and Oryza sativa with those from maize genome sequence to mine novel SBP TFs in maize. A total of 37 SBP TFs distributed in eight chromosomes were identified. Phylogenetic analysis indicated that SBP transcription factor genes have stronger homology, especially between Zea mays and Oryza sativa. Moreover, Promoters-cis Elements analysis of those SBP TFs demonstrated that they might be involved in plant growth and development, morphogenesis, adversity response, the development of flower organs and photosynthesis. It is probable that SBP TFs regulate plant growth and development through the multiple hormone of signaling transduction pathway, such as gibberellin, auxin, abscisic acid, and salicylic acid.

GhHMGR Gene Function in Ovule Development of Cotton (Gossypium hursutum L.)

MA Fu-Lei,LI De-Mou,LI Zhi,YANG Wei-Juan,ZHOU Xue,YOU Yu,LUO Xiao-Ying

Acta Agron Sin. 2016, 42(02):  222-229.  doi:10.3724/SP.J.1006.2016.00222

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The constitutive promoter CaMV35S was selected to drive the expression of GhHMGR gene in cotton by using the plant genetic engineering technology. The HMGR contents of ovule (10 DPA) of transgenic lines were detected, and the contents of total sugar, oil and protein as well. Meanwhile, we also performed ovule culture in vitro. The results showed that the expression level of GhHMGR was higher in part organ under light, such as petiole and boll shell than in root and ovule in dark. The GhHMGR gene could partly recover Arabidopsis (Arabidopsis thaliana) hmgr mutant characteristics. Compared with wild-type（WT）, the HMGR contents of ovule (10 DPA) in overexpression transgenic lines increased, whereas decreased in suppression expression lines. While the contents of total oil and protein of overexpression transgenic lines increased, and that of total sugar decreased. Meanwhile, the suppression expression lines performed in an opposite trend. The investigation of ovule culture in vitro showed that ovules became deformity after treated with the inhibitor. All of these implied that expression of GhHMGR gene plays a key role in the development of cotton ovule.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

Effect of Alternate Irrigation in Partitioned Roots on the Kernel-filling and Its Related Physiological Characteristics in Maize

XU Yun-Ji,QIAN Xi-Yang,LI Yin-Yin,WANG Zhi-Qin,YANG Jian-Chang*

Acta Agron Sin. 2016, 42(02):  230-242.  doi:10.3724/SP.J.1006.2016.00230

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This study investigated whether and how post-tasseling alternate irrigation in partitioned roots could enhance the filling of inferior caryopses in maize. A high-yielding maize cultivar, Denghai 11, was grown in a glasshouse, and two irrigation treatments, conventional irrigation (CI) and alternate irrigation in partitioned roots (PAI), were applied from tasseling to maturity. Kernel filling rates and starch accumulation rates of superior and inferior caryopses and changes in ethylene evolution rate and polyamine contents in caryopses, photosynthetic and senescence characteristics of the ear leaf and non-structural carbohydrates (NSC) in stems were determined. The results showed that, compared with CI, PAI significantly increased kernel yield, strengthened photosynthetic characteristics of the ear leaf during late kernel-filling period, delayed leaf senescence, promoted NSC remobilization from stems, and increased free-spermidine (free-Spd) and free-spermine (free-Spm) contents, whereas declined the free-putrescine (free-Put) content and ethylene evolution rate in inferior caryopses. No significant effect of PAI on the kernel filling of superior caryopses was detected. Correlation analysis showed that kernel-filling rates and starch accumulation rates were very significantly and positively correlated with free-Spd and free-Spm contents, and significantly and negatively correlated with ethylene evolution rate. The results indicate that PAI enhances the filling of inferior caryopses and increases kernel yield through increasing photosynthetic ability of the ear leaf, remobilization of NSC from stems, and free-Spd and free-Spm contents, and decreasing ethylene evolution rate in inferior caryopses during the grain-filling period.

Comparative Proteomics Analysis of R. glutinosa Tuber Root in Response to Consecutive Monoculture

WU Lin-Kun,CHEN Jun,WU Hong-Miao,WANG Juan-Ying,QIN Xian-Jin,ZHANG Zhong-Yi,LIN Wen-Xiong

Acta Agron Sin. 2016, 42(02):  243-254.  doi:10.3724/SP.J.1006.2016.00243

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Rehmannia glutinosa L. is widely applied in Chinese medicine. However, consecutive monocropping of this plant results in serious decline in both biomass and quality of underground tubers, with poor field performance and insufficient resistance to disease and pest. In the present study, the tuber roots from the newly planted (NP) and consecutively monocropped (SM) R. glutinosa were used through comparative proteomics analysis to study the response of R. glutinosa to consecutive monocropping and the underlying mechanisms of replanting disease. Comparative proteomics analysis showed that these proteins involved in important physiological processes and biosynthese of main components in tuber roots were significantly down-regulated under consecutive monocropping regime.It was also found that chaperonins related to protein folding was down-expressed with the extended monocropping. However, these proteins related to stress response/defense such as pathogenesis-related protein 10, cytochrome P450 and Type IIIa membrane protein cp-wap13 were up-regulated in consecutively monocropped R. glutinosa. Quantitative analysis by qRT-PCR confirmed the up-regulation of PR-10 responding to consecutive monocropping or the infection of pathogenic Fusarium oxysporum. Moreover, PR-10 was gradually up-regulated with the increasing days of infection. In conclusion, consecutive monocropping of R. glutinosa greatly affects the expression profile of proteome in tuber roots. These abnormally expressed proteins might lead to the metabolic disturbances and low energy production. Moreover, the limited energy is applied to resist the external environmental stresses, resulting in significant decline in the growth of tuber roots and the accumulation of active ingredients.

Effects of 1,2,4-Trichlorobenzene on Photosynthetic Characteristics of Flag Leaf during Grain Filling Stage and Grain Yield of Two Rice Cultivars

LI Yu,CHEN Lu,YAN Kai,SUN Ying,YIN Yi-Fan,DING Xiu-Wen,DAI Qi-Gen,ZHANG Hong-Cheng

Acta Agron Sin. 2016, 42(02):  255-264.  doi:10.3724/SP.J.1006.2016.00255

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A pot experiment was conducted, using two rice cultivars Ningjing 1 (TCB sensitive) and Yangfujing 8 (TCB tolerant), with five concentration treatments (0, 10, 20, 40, and 80 mg TCB per kg dry soil) to explore the responses of two rice cultivars to TCB, and provide the basis for the high, stable, and safe production of rice. The results indicated that significant differences were found in the effects of TCB treatments on grain yield and photosynthetic parameters of flag leaf between the two cultivars. Plant height, fresh weight, and yield were significantly increased, chlorophyll content, net photosynthetic rate, intercellular CO₂ concentration, transpiration rate, and q_n were slightly increased, while F_v/F_m and F_v/F_o were slightly decreased in the low TCB concentration (10 mg kg^-1) treatment of Yangfujing 8. Under the same condition, the net photosynthetic rate, CO₂ intercellular concentration, transpiration rate, Φ_PSII, F_v/F_m, F_v/F_o, q_p, yield in Ningjing 1 were slightly declined with significant reduction of stomatal conductance. The photosynthetic characteristics, yield, plant height, fresh weight in Ningjing 1 were decreased significantly, while Yangfujing 8 showed more resistance and adaptation to TCB at 20 mg kg^-1. Both cultivars showed significant decrease in growth, photosynthesis and yield in treatments with high TCB concentrations (40 and 80 mg kg^-1), with the greater decrements in Ningjing 1. The effect of TCB on photosynthetic characteristics and yield of rice was not only related to TCB concentration, but also to cultivars. Low TCB levels slightly promoted the plant height, fresh weight, chlorophyll content, photosynthetic characteristics and rice yield in Yangfujing 8 that showed stronger tolerance to TCB than Ningjing 1, under high TCB concentrations (40 and 80 mg kg^-1). 

Dynamic Model and Its Characteristics Analysis for Dry Matter Production after Heading of Indica/JaponicaHybrid Rice of Yongyou Series

WEI Huan-He,MENG Tian-Yao,LI Chao,ZHANG Hong-Cheng,SHI Tian-Yu,MA Rong-Rong,WANG Xiao-Yan,YANG Jun-Wen,DAI Qi-Gen,HUO Zhong-Yang,XU Ke,WEI Hai-Yan,GUO Bao-Wei

Acta Agron Sin. 2016, 42(02):  265-277.  doi:10.3724/SP.J.1006.2016.00265

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This study was conducted using indica-japonicahybrid rice Yongyou 1538 and Yongyou 7176 as the material, conventional japonica rice Ningjing 3 and Wuyunjing 24, and hybrid indica rice Yangliangyou 6 and Liangyoupeijiu as the check to study the characteristics of dry matter production after heading inindica/japonica hybrid rice of Yongyou series and compare the differences in characteristics of dry matter production after heading among different types of rice variety. Results indicated that, grain yield of indica/japonica hybrid rice of Yongyou series was 11.5 t ha^-1, on an average, which was 7.8% and 10.4% higher than that of conventional japonica rice and hybrid indica rice, respectively. Dry matter accumulation of indica/japonica hybrid rice was 8.9 t ha^-1, which was19.1% and 26.9% higher than that of conventional japonica rice and hybrid indica rice, respectively.Richards' equation was fit to simulate the relationship between dry matter weight and days after heading for three types of rice variety (R²≥0.990). Rate of dry matter accumulation of three types of rice variety decreased after an even increase.Maximum rate of biomass accumulation and mean rate of biomass accumulation after heading of hybrid indica rice were the highest, followed by those of conventional japonica rice, and indica/japonica hybrid rice. Days to maximum rate of dry matter accumulation after heading of indica/japonica hybrid rice was at 42–44 d after anthesis, while those of conventional japonica rice and hybrid indica rice were both at 26–28 d after anthesis. Duration of gradual increase stage and dry matter accumulation in this stage were higher in indica/japonica hybrid rice than inthe check, while mean rate of dry matter accumulation during gradual increase stage was higher inhybrid indica rice than inindica/japonica hybrid rice. The duration and dry matter accumulation during fast increase stage and slow increase stage of conventional japonica rice were the highest among three types of rice variety, while mean rate of dry matter accumulation during fast increase stage and slow increase stage of hybrid indica rice was the highest. Our results implied that greater dry matter accumulation mainly occurred in the gradual increase stage for indica/japonica hybrid rice, which was mainly attributed to the longer duration of this stage.

Microstructure of Glandular Trichomes on Leaf Surface of Sesame and Changes of Trichome Secretions under Drought Condition

SU Shi,LI Rui-Hang,LANG Dan-Ying,ZHANG Ke,HAO Xiao-Hu,LIU Yan,WANG Jun-Wei,XU Hong

Acta Agron Sin. 2016, 42(02):  278-294.  doi:10.3724/SP.J.1006.2016.00278

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We investigated the microstructure of glandular trichome on sesame leaf surface by scan electron microscope and extracted the trichome secretions using dichloromethane as solvent, and the secretion components under normal and drought conditions were measured by GC/MS to compare the secondary metabolites between sesame varieties. The experiment results indicated that, there were non-glandular hairs, long stalk glandular hairs, short stalk glandular hairs and mucilage hairs on the sesame leaves surface, and the stomata belong to the buttercup type. The secretions of glandular hairs contained a variety of components, of which the main fractions with peak area more than 2% were composed of esters and straight chain saturated alkanes mainly, showing notable differences among four sesame cultivars. Under drought condition, the secreted components changed significantly. Ji9014 with the highest drought resistance among the four cultivars, had ten components, including C36 alkanes and C34 alkanes, which were higher in Ji9014 than in other tested cultivars. We can conclude that, the GC/MS technique is feasible to identify the glandular trichome secretions, and the secretion components are good index to reveal differences between varieties and the effects of drought condition on metabolic pathway in sesame leaves. So, the glandular hairs and secretion components have important application values in sesame germplasm identification and molecular breeding for drought-resistant sesame.

RESEARCH NOTES

Expression Analysis of Two CIPK genes under Abiotic Stress in Foxtail Millet

U Ai-Li,ZHAO Jin-Feng,WANG Gao-Hong,DU Yan-Wei,LI Yan-Fang,ZHANG Zheng,GUO Er-Hu,LIANG Ai-Hua

Acta Agron Sin. 2016, 42(02):  295-302.  doi:10.3724/SP.J.1006.2016.00295

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CIPK (CBL interacting protein kinase) is a type of serine or threonine protein kinases, which plays an important role in response to stress. In this study, we identified two CIPK genes designated as SiCIPK6 and SiCIPK16 from foxtail millet (Setaria italica) genome using bioinformatics methods. The sequence analysis showed that SiCIPK6 has a length of 1994 bp in the genome, encoding 513 amino acids residues, and SiCIPK16 is 1885 bp, encoding 473 amino acids residues. These two genes have no alternative splicing and intron. The characters predicted based on the bioinformatics analysis revealed that the protein sequences and structure of the two SiCIPK genes were very conservative just like CIPK genes in other species. Real-time PCR analysis discovered that the expression of SiCIPK6 and SiCIPK16 was up-regulated by ABA, cold, heat, drought and salt stress, respectively. The expression was strongly induced by ABA, drought and salt treatments for SiCIPK6, and by cold, drought and heat treatments for SiCIPK16. The semi-quantitative PCR analysis showed that SiCIPK6 and SiCIPK16 were expressed at the jointing, booting and filling stages, and induced by drought stress in the corresponding growth period. Foxtail millet CIPK genes reported in this study would enrich CIPK members in plant kingdom and provides important information for further elucidating the function and mechanisms of the CBL/CIPK network system responsing to stresses in foxtail millet.

Analysis of QTL for Fatty Acid Contents under Different Environments in Soybean

LEI Ya-Kun,LIU Bing-Qiang,DI Rui,YAN Long,YANG Chun-Yan,HAO Dong-Xu,ZHANG Meng-Chen

Acta Agron Sin. 2016, 42(02):  303-310.  doi:10.3724/SP.J.1006.2016.00303

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Soybean is an important crop which contributes about 60% of the world’s oilseed production. The quality of soybean oil depends on the relative composition of fatty acid in seeds. A population of recombinant inbred lines  derived from a cross of Jidou 12 × Heidou was grown in two environments and the seed samples from the environments were evaluated for fatty acid contents. Simple sequence repeat (SSR) markers were used to construct genetic linkage map. A total of 16 QTLs underlying fatty acid contents were identified by CIM and MCIM methods using Windows QTL cartographer 2.5 and QTL Network-2.0 software at Sanya and Shijiazhuang locations, respectively. These QTLs were scattered on linkage groups A2, B2, C2, F, G, I, L. According to the two environment combined data, 13 QTLs were detected using two mapping methods. Nine of them were common in the results from two methods. Another QTL associated with stearic acid content, named as Ste-1, located on LG B2 and flanked by Satt168 and Satt556, was stable across two locations, also. QTL Ste-1 could explain 12% of the phenotypic variation at both locations. This study is helpful to improve fatty acid composition in soybean.