Table of Content

12 April 2017, Volume 43 Issue 04

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Identification and Gene Mapping of Starch Accumulation and Early Senescence Leaf Mutant esl9 in Rice

XIAO Yan-Hua**, CHEN Xin-Long**, DU Dan, XING Ya-Di, ZHANG Tian-Quan, ZHU Mao-Di,LIU Ming-Ming,ZHU Xiao-Yan, SANG Xian-Chun,HE Guang-Hua*

Acta Agron Sin. 2017, 43(04):  473-482.  doi:0.3724/SP.J.1006.2017.00473

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A new leaf senescence mutant esl9(early senescence leaf9) was discovered from the progeny of indica maintainer line 1B mutatedby ethylmethanesulfonate(EMS). Compared with the wild-type, the leaf of esl9 was pale green at seedling stage; chlorosis occurredatleaf tipand gradually extendedto the middle-upper parts of leaf at tillering stage. However, the leaf base remained green untilmaturity. In theesl9, the photosynthetic pigment contentsdeclined,and the contentsof reactive oxygen species (ROS), such as, O₂^‾, ·OH and H₂O₂, heightened compared with those in the wild-type. At the same time, activities of protective enzymes, SOD and CAT, both reduced in esl9. The results of iodine-iodide kalium dyeing and starch content determinationshowed that more starch granules accumulated in the esl9 leaf. Quantitative RT-PCR results indicatedthat genes responsible for starch synthesis were up-regulated and genes participated in the triose phosphatedistribution path were down-regulated. We made an inferencethatgene mutation changed the distribution of triose phosphate, resulting in starch granules accumulating in the leaf, chloroplast structure being destroyed and photosynthetic systembeing blocked, thus increasingthe contents of ROS,eventually causing leaf senescence.Genetic analysis demonstrated that the phenotype of esl9 was controlled by a dominant nuclear gene. The target geneESL9 was mapped between SSR markers S11-110 and S11-87 with a physical distance of 304.9 kbon chromosome 11. These results will lay a foundation for cloning and functionallyanalysing ESL9.

Linkage Analysis and Genome-Wide Association Study of QTLs Controlling Stem-Breaking-Strength-Related Traits in Wheat

LIU Kai,DENG Zhi-Ying,ZHANG Ying,WANG Fang-Fang,LIU Tong-Tong,LI Qing-Fang,SHAO Wen,ZHAO Bin,TIAN Ji-Chun*,CHEN Jian-Sheng*

Acta Agron Sin. 2017, 43(04):  483-495.  doi:10.3724/SP.J.1006.2017.00483

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Stem strengthhas close relationship with lodging character, thereby, affects final yield in wheat. The objectives of this study were to unravel the genetic mechanism of stem-breaking-strength-related traits and find molecular markers closely linked or associated with these traits. We tried to map the stem-breaking-strength-related QTLs through linkage analysis using the RIL population consisting of 173 F_8:9 lines derived from Shannong 01-35 ´ Gaocheng 9411) and associationanalysis using a nature population consisting of 2015 wheat varieties. Both populations were planted in two environments and genetically screened with the 90 k SNP array, DArT technology, and traditional molecular markers.By means of the existing high-density genetic map, nine additive QTLs were detectedin different regions onchromosome 4B, such asDURUM_CONTIG63670_287–IACX557 and EX_C101685–RAC875_C27536, which controlled stem-breaking strength, plant height, filling degree of the second internode and culm wall thickness of the second internode and explained 9.40–36.30% of the phenotypic variations.By means of a composite map (containing 24,355 SNPs) based on the IlluminaInfinium assay, a total of 37 SNPs were found in the natural population to be associated withstem-breaking-strength-relatedtraits (P<0.0001). These SNPswere distributed on chromosomes1B, 2B, 2D, 3A, 3B, 4B, 5A, 5D, 6B, 7A, 7B, and 7D and explained 10.70%–36.30% of the phenotypic variations.Thegenetic distance between RAC875_C27536 detected throughlinkage analysis and Tdurum_contig4974_355 detected through genome-wide association analysis was 6.7cM in thecomposite map, indicatingthe presence of importantgenes controlling stem strengthin this region.

Mapping QTLs for Awn Length in Recombinant Inbred Line Population Derived from the Cross between Common Wheat and Tibetan Semi-wild Wheat

GONG Xi,JIANG Yun-Feng,XU Bin-Jie,QIAO Yuan-Yuan,HUA Shi-Yu,WU Wang,MA Jian,ZHOU Xiao-Hong,QI Peng-Fei,LAN Xiu-Jin

Acta Agron Sin. 2017, 43(04):  496-500.  doi:10.3724/SP.J.1006.2017.00496

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Awn length is an important agronomic trait in hexaploid wheat and controlled by multiple genes. A recombinant inbred line (RIL) population containing 186 lines were developed by crossing Tibetan semi-wild wheat accession Q1028 (female) and common wheat variety Zhengmai 9023, and a genome-wide genetic map (2597 cM) was constructed using SSR and DArT markers. QTLs controlling awn length were identified with two-year phenotypic data and the genetic map by using the Inclusive Composite Interval Mapping (ICIM) method. Two QTLs associated with awn length, designated Qwa.sau-4AS and Qwa.sau-5AL, were detected on chromosomes 4A and 5A, which explained 7.4% and 27.3% of phenotypic variations, respectively. According to genetic locations, effects of the two QTLs might be from Hd and B1 genes that could reduce awn length, respectively. Genetic analysis showed that Qwa.sau-4AS and Qwa.sau-5AL had cumulative effect, with a stronger inhibiting effect in Qwa.sau-5AL than in Qwa.sau-4AS. These results are valuable for fine mapping and cloning target genes controlling awn length in the future.

Quality Characteristics of Winter Wheat Varieties Tested in National Trials in Northern Region and Yellow-Huai River Valley Winter Wheat Zone from 2000 to 2015

HU Xue-Xu,SUN Li-Juan,ZHOU Gui-Ying,WU Li-Na,LU Wei,LI Wei-Xi,WANG Shuang,YANG Xiu-Lan,SONG Jing-Ke,WANG Bu-Jun

Acta Agron Sin. 2017, 43(04):  501-509.  doi: 10.3724/SP.J.1006.2017.00501

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Wheat varieties tested in national regional trials may be released as major cultivars in wheat production in the near future. Thus, the quality level and distribution character of these varieties can be a hint to guide wheat quality breeding. A total of 1589 samples of 1001 varieties were collected from the regional trails in Northern Region and Yellow-Huai River Valley Winter Wheat Zone form 2000 to 2015, and classified into strong-gluten, medium-strong-gluten, and medium-gluten types according to the national standards of wheat quality. Eight indices were measured to analyze the changing trend of winter wheat quality of newly developed and registered varieties. In general terms, the quantity of wheat varieties tested increased year by year while the quality of varieties was desired to be improved. The ratio of registered varieties showed a declined trend from 2000 to 2015, particularly, for the medium-gluten type. In contrast, the proportion of medium-strong-gluten varieties increased in the registered varieties. The medium-gluten varieties had high contents of protein and wet gluten, and moderate levels of sedimentation volume, dough stability time, extension area, and maximum resistance. There were significant difference among quality types in sedimentation value, dough stability time, extension area, and maximum resistance (P < 0.05), with remarkable variations in all quality indices across years, showing an order of strong-gluten type > medium-strong-gluten type > medium-gluten type. In the strong-gluten varieties, the protein quality indices were significantly lower than those in the control varieties. Our results indicate that wheat quality breeding has achieved progresses in China in recent 16 years, but the overall quality indices of varieties should be further improved, especially for the strong-gluten varieties. More medium-strong-gluten varieties should be employed in Northern Region and Yellow-Huai River Valley Winter Wheat Zone in the future 5–10 years.

Molecular Cloning and Functional Identification of Peroxidase Gene ScPOD02 in Sugarcane

SU Ya-Chun,WANG Zhu-Qing,LI Zhu,LIU Feng,XU Li-Ping*,QUE You-Xiong,DAI Ming-Jian,Chen Yun-Hao

Acta Agron Sin. 2017, 43(04):  510-521.  doi:10.3724/SP.J.1006.2017.00510

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Peroxidases (PODs), widely existing in various plant organs and different development stages, play a vital role in plant growth and development, and also respond to adverse stresses. Based on the previous transcriptome data, we isolated a cDNA (GenBank Acc. No. KU593507) and genomic DNA (GenBank No. KU593508) sequences of ScPOD02 from smut resistant genotype Yacheng 05-179 infected by Sporisorium scitamineum for two days. Sequence analysis showed that the full length cDNA of ScPOD02 is 1434 bp with an ORF of 1047 bp in length, encoding 348 amino acids. Its genomic DNA length is 1558 bp containing two exons and one intron. Phylogenetic tree analysis demonstrated that ScPOD02 and rice OsPrx11 (GenBank Acc. No. gi|55700889) were clustered into the same evolutionary branch, suggesting that ScPOD02 belongs to one of the acidic exocytosis/cell wall type of class I.1 peroxidase family members. ScPOD02 was further ligated with a prokaryotic expression vector of pET 32a and then transformed into Escherichia coli BL21. An approximately 60 kD fusion protein was obtained by isopropyl-β-d-thiogalactoside induction. Compared with the control, the growth of recombinant BL21 cells was enhanced under the stress of polyethylene glycol, indicating its high tolerance to drought stress. RT-qPCR analysis showed that the transcripts of ScPOD02 were up-regulated in sugarcane smut-resistant cultivars (YZ03-258, YZ01-1413, YT96-86, and LC05-136) by S. scitamineum except for ROC22 and YZ03-103, but remained or slightly decreased in the middle-susceptible (FN39 and GT02-467) and susceptible (FN40) cultivars. In addition, ScPOD02 positively responded to salicylic acid, abscisic acid, polyethylene glycol and sodium chloride stresses. The transient expression of ScPOD02 in Nicotiana benthamiana was performed using Agrobacterium mediated method. A deeper DAB staining color in N. benthamiana leaves was observed after overexpressing ScPOD02. Furthermore, the target gene ScPOD02 and the N. benthamiana hypersensitive reaction (HR) marker genes (NtHSR201 and NtHSR203) and ethylene synthesis dependent genes (NtEFE26 and NtAccdeaminase) were all up-regulated. These reach a conclusion that the ScPOD02 has potential roles in the immune response and in protecting sugarcane from drought and salt stresses.

Genetic Analysis and Gene Mapping of Yellow leaf and dwarf (yld) mutant in Rice

LI Zi-Zhuang,XU Qian-Kun,YU Hai-Ping,ZHOU Ting-Ting,XUE Da-Wei,ZENG Da-Li,GUO Long-Biao,QIAN Qian,REN De-Yong

Acta Agron Sin. 2017, 43(04):  522-529.  doi:10.3724/SP.J.1006.2017.00522

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Leaf color mutants of rice are ideal materials in studies on photosynthesis, chlorophyll metabolism and chloroplast development in plants. A yellow leaf and dwarf mutant yldwas obtained from ethyl methane sulfonate (EMS)-treated Shuhui 527 (Oryza sativa L.). Compared with the wild type, the yld mutant showed yellow leaf and dwarfism, and the contents of chlorophyll and carotenoid were obviously decreased. Transmission electron microscope observation revealed that the structure of most chloroplasts seemed to be normal, however, with the fuzzy grana, and fewer and looser stroma lamella in the yld mutant. Meantime, plant height, branch number, grain number per panicle, 1000-grain weight and seed-setting rate were significantly decreased, while the number of effective panicle was obviously increased in the ylda single recessive gene. The 323 mutational individuals from the F₂ generation of the cross of Wuyunjing 7 and yld mutant were used for gene mapping. Finally, the YLD locus was mapped on chromosome 11 between two Indel markers L5 and L7, with an approximate 115 kb physical region. This result would facilitate cloning and functional analysis for the YLD gene. mutant compared with those in the wild type. Genetic analysis showed that the yld mutant traits were controlled by

Analysis of Genetic Diversity in Common Millet (Panicum miliaceum) Using Fluorescent SSR in China

WANG Rui-Yun,JI Xu,LU Ping,LIU Min-Xuan,XU Yue,WANG Lun,WANG Hai-Gang,QIAO Zhi-Jun

Acta Agron Sin. 2017, 43(04):  530-548.  doi:10.3724/SP.J.1006.2017.00530

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Evaluating the genetic diversity of germplasm resources in common millet is helpful to facilitate the understanding of its origin and evolution, and facilitate to explore the elite germplasm for highly effective utilization. Genetic diversity among 132 accessions of common millet from 11 provinces in China was detected by 15 millet-specific fluorescent-labelled simple sequence repeat (SSR) primers. A total of 107 alleles were detected, with alleles per locus ranging from 2 to 14 (mean = 7). Polymorphism information content and expected heterozygosity ranged from 0.0893 to 0.8538 (mean = 0.4864) and from 0.0936 to 0.8676 (mean=0.5298), respectively. Cluster analysis based on genetic distance separated the accessions into four groups, including Group I from Northeast spring-sowing ecotope, Group II from Loess Plateau spring & summer-sowing ecotope, Group III from North spring-sowing ecotope, and Group IV from North spring-sowing ecotope and Loess Plateau spring & summer-sowing ecotope. Model-based genetic structure analysis indicated that common millet accessions from China were derived from four (Northeast, Loess Plateau, North and Northwest) gene pools. The above two cluster analyses uncovered a close correlation between geographical regions and genetic diversity. There were abundant genetic variations from different accessions of common millet. The obtained information would provide an accurate estimation of the genetic diversity of common millet in China at molecular level.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

Application of an Improved Method in Retrieving Leaf Area Index Combined Spectral Index with PLSR in Hyperspectral Data Generated by Unmanned Aerial Vehicle Snapshot Camera

GAO Lin,YANG Gui-Jun,LI Chang-Chun,FENG Hai-Kuan,XU Bo,WANG Lei,DONG Jin-Hui,FU Kui

Acta Agron Sin. 2017, 43(04):  549-557.  doi:10.3724/SP.J.1006.2017.00549

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The objective of this study was to demonstrate the value of an improved method of retrieved leaf area index (LAI) based on unmanned aerial vehicle (UAV) hyperspectral data combined spectral characteristics, as red edge parameters (REPs) and vegetation indices, with partial least squares regression (PLSR). We got UAV UHD185 hyperspectral images at booting, anthesis, and filling stages in winter wheat. And synchronously measured ASD hyperspectral data and winter wheat LAI. We compared UHD185 data with ASD data in terms of the correlation between reflectivity and vegetation indices to verify the UAV hyperspectral data accuracy. The band 3 to 96 (458-830 nm) of UHD185 hyperspectral data had better spectral quality and was suitable for detecting winter wheat LAI. We did correlation analysis between spectral characteristics, six kinds of vegetation indices and four kinds of red edge parameters, and LAI, and used two kinds of validation methods, independent validation and cross validation, to analyze the prediction accuracy of winter wheat LAI. Compared with traditional LAI fitting method, the improved LAI fitting method especially PLSR+REPs, greatly improved the prediction accuracy of winter wheat LAI. The above results confirmed that the improved LAI fitting method is able to better utilize UAV UHD185 hyperspectral data to predict LAI of winter wheat. Moreover, it is expected to provide a few new ideas for retrieving crop physical and chemical parameters based on UAV hyperspectral data.

Potassium Absorption and Utilization Characteristics of Rice Varieties with the Highest Population Productivity under corresponding Nitrogen Fertilization in Huaibei Area

LIANG Jian,REN Hong-Ru,XIA Min,LI Xiao-Feng,CHEN Meng-Yun,LI Jun,ZHANG Hong-Cheng,HUO Zhong-Yang*?

Acta Agron Sin. 2017, 43(04):  558-570.  doi:10.3724/SP.J.1006.2017.00558

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A field experiment was carried out using 34 medium-maturing medium japonica rice varieties grown in Huaibei area with seven nitrogen application levels (0, 150.0, 187.5, 225.0, 262.5, 300.0, and 337.5 kg ha–1) to investigate the relationship between potassium and yield. According to the highest population productivity under corresponding N fertilization, rice varieties were classified into four types including top type (TT), high type (HT), middle type (MT), and low type (LT). Yield components, and K absorption and translocation of the four types of rice variety were compared. K concentration of aboveground parts of plant steam-sheath and leaf in different types was the highest at heading stage and that was higher in TT than the other three types. K concentration of panicle was no significant difference among tested varieties. With increasing productivity level, total K accumulation increased. From transplanting to jointing stage, the K accumulation in LT was 120.56 kg ha-1, accounting for 50.56%. The K accumulation of TT was 108.02 kg ha-1, accounting for 35.99%. After jointing, the K accumulation and ratio at each growth stage of TT were higher than those of other three types. The K uptake rate showed a trend of MT > LT > HT > TT from transplanting to jointing stage, and TT > HT > MT > LT from jointing to heading stage and from heading to maturity stage. K translocation and K translocation efficiency of leaf were obviously higher than those of steam-sheath. K translocation of TT and HT’s K translocation efficiency were both the highest. The increasing in K of panicle increased with increasing productivity level. From heading to maturity stage, K translocation conversion rate of vegetative organ was the highest in LT, medium in MT, and the lowest in TT. Internal nutrient efficiency and K requirement for 100 kg grain among four types were no obvious difference and K partial factor productivity and harvest index of K showed a trend of TT > HT > MT > LT. In conclusion, the higher the production level, the higher the potassium use efficiency of plants and organs at the middle and later periods of development. Maintaining high K uptake and translocation efficiency after heading is an important characteristic of high-yield rice varieties.

Effects of Mulch Farming with Different Films on Weed Infestation, Soil Moisture and Temperature, as well as Yield and Quality of Potato

ZHANG Shu-Min,NING Tang-Yuan,LIU Zhen,WANG Bin,SUN Tao,ZHANG Xue-Peng,HE Zhen-Kun,YANG Yan,MI Qing-Hua

Acta Agron Sin. 2017, 43(04):  571-580.  doi:10.3724/SP.J.1006.2017.00571

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Plastic film mulching is widely used on potato cultivation, however, leads to a serious “white pollution” and weed infestation. Three types of films including black-white match color film (BW), biodegradable film (BI), and common plastic film (CK), were used on mulching potato variety Helan 15 from 2013 to 2015, to study their effects on weed infestation, soil moisture and temperature, and yield and quality of potato in North China. BW and BI significantly decreased weeds densities by 44.1% to 58.5% and 26.7% to 38.3% compared with CK, respectively. CK had a higher warming effect than BW and BI during the earlier stages of potato growth, while BW had relatively small variations in daily temperature at the starch accumulation stage. Compared with CK and BI, BW treatment stored more soil moisture in the dry year (2014). Compared with CK, yields of BW in the normal (2013) and dry year (2014) increased by 6.2% and 8.2% respectively, and that of BI increased by 7.1% in dry year. However, there was no significant difference in yield between treatments in the wet year (2015). Compared with CK, the starch content in tubers of BW and BI increased by 18.3% to 37.6% and 7.0% to 28.9%, respectively; the vitamin C content of BW increased by 1.0% to 4.3%; however, the protein contents of BW and BI were lower. Therefore, mulch farming with black-white match color film and biodegradable film instead of common plastic film can decrease weeds densities and create suitable soil temperature and moisture conditions for improving yield and quality of potato.

Effect of Mechanical Transplanting with Pothole Seedlings on Grain Quality of Different Types of Rice in Rice-Wheat Rotation System

XING Zhi-Peng,ZHU Ming, WU Pei,QIAN Hai-Jun,CAO Wei-Wei,HU Ya-Jie,GUO Bao-Wei,WEI Hai-Yan,XU Ke,HUO Zhong-Yang,DAI Qi-Gen,ZHANG Hong-Cheng*

Acta Agron Sin. 2017, 43(04):  581-595.  doi:10.3724/SP.J.1006.2017.00581

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In rice-wheat rotation system, the mechanical transplanting with pothole seedlings (MT) is a new developed high yield and high efficiency rice planting method with increased promotion area year by year. Understanding the rice quality traits under MT method is of great importance to study and develop high yield and high quality cultivation techniques under mechanical conditions in rice-wheat rotation system. The comparison experiments were carried out to investigate the difference in processing, appearance, nutrition, cooking and eating, and taste quality, and starch RVA profile of conventional japonica, indica hybrid and japonica-indica hybrid rice, by using MT, mechanical transplanting with carpet seedlings (MC) and mechanical direct seeding (MD) in high-yield cultivation condition, which would provide theoretical reference and practice data for high yield and high quality rice production. The head rice rate and head rice yield under MT were 1.6% and 7.8% larger than those under MC, and 4.9% and 25.9% larger than those under MD (P<0.05), respectively, indicating a better processing quality under MT method. Compared with MC and MD, MT decreased appearance quality with increased chalky grain rate, chalkiness size and chalkiness degree (P<0.05), and increased protein content and nutrition quality with the increase of 8.7% and 28.0% in protein yield, respectively. The effects on amylose content, setback and pasting temperature were MTMC>MD, which indicated that the cooking and eating quality under MT method might be meliorated. The taste quality was no significant difference among planting methods, showing a tendency of MTMT>MD for indica hybrid rice. Therefore the MT method is recommended as one of promising mechanized planting methods to simultaneously realize high yield, high efficiency and high quality in rice production of rice-wheat rotation system.

Effect of Balanced Fertilizer Application on Crop Yield in Potato-Soybean Relay-Cropping System

CHEN Guang-Rong,WANG Li-Ming,YANG Ru-Ping,DONG Bo,YANG Gui-Fang,ZHANG Guo-Hong,YANG Wen-Yu

Acta Agron Sin. 2017, 43(04):  596-607.  doi:10.3724/SP.J.1006.2017.00596

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The treatments of optimum fertilization (OPT:N 180, P 135, K 135 kg ha–1), OPT-N, OPT-P, OPT-K, OPT-1/3N, OPT +1/3N, OPT-1/3P, OPT +1/3P, OPT-1/3K, and CK were designed according to soil condition and uptake characteristics of nitrogen, phosphorus and potassium of potato/soybean relay-cropping system in Northwest irrigation districts. A field experiment was conducted in two consecutive seasons (from 2012 to 2013) to verify the yield response to balanced fertilization. The relay-cropping potato yield of OPT was significantly higher than that of OPT-N, OPT-P, and OPT-K, with obvious difference of 11 653.86 kg ha–1 between OPT and OPT-N, the relay-cropping soybean yield of OPT was significantly higher than that of OPT-N, OPT-P, and OPT-K, with obvious difference of 751.55 kg ha–1 between OPT and OPT-P. Therefore, nitrogen and phosphorus were the first limiting factors in relay-cropping potato and soybean production. Under certain phosphorus and potassium fertilizer, the relay-cropping potato yield increased with increasing N application, reaching the highest of 50 231.85 kg ha–1 under 240 kg ha–1 treatment (OPT+1/3N), but there was no significant difference between OPT and OPT+1/3N. And the relay-cropping soybean yield increased firstly and decreased then with increasing nitrogen fertilizer application, with the highest yield of 3373.55 kg ha–1 under 180 kg ha–1 treatment (OPT), there was no significant difference between OPT, OPT-1/3N, and OPT+1/3N. Under certain nitrogen and potassium fertilizer, the yield of relay-cropping potato and soybean increased with the increase of phosphorus fertilizer application, and reaching the highest yield of 52 430.03 kg ha–1 and 3637.13 kg ha–1 under 180 kg ha–1 treatment (OPT+1/3P), respectively. For relay-cropping potato, average fresh-weight of individual tuber reached the highest under 180 kg ha–1 treatment (OPT+1/3P), which was 185.13 g. For relay-cropping soybean, the average of pods per plant, seeds per plant and seeds per pod reached the highest under OPT+1/3P, which were 74.24, 1.87, and 139.15 respectively. Therefore, nitrogen (N 180 kg ha–1) fertilizer was adequate, phosphorus (P 135 kg ha–1) fertilizer a little lacking, and potassium (K 135 kg ha–1) fertilizer on the high side in OPT treatment.

RESEARCH NOTES

Differential Expression Characteristics of Proteins Involved in Early Development of Maize Grain

YU Tao,LI Geng,ZHANG Cheng-Fen,LIU Peng,DONG Shu-Ting,ZHANG Ji-Wang,ZHAO Bin

Acta Agron Sin. 2017, 43(04):  608-619.  doi:10.3724/SP.J.1006.2017.00608

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During early stage of maize grain development, the metabolic activity is strong and the cell division and enlargement processes are also active, leading to increase the grain sink size for subsequent accumulation of storage material. To explore the protein synthesis, accumulation and regulation during early maize grain development, grains of maize cultivar Denghai 661 in the middle of ear were harvested at 3, 5, and 10 days after flowering artificial saturation pollination, respectively, and analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) proteomics. A total of 2639 proteins were identified and quantified in maize grain during early stages of development, showing that these proteins were involved in diverse biological processes and molecular functions, of which the metabolic process and molecular processes were the two most important biological processes, and the catalytic activity and binding were the two most important molecular functions, all of them played important roles in maize grain development. Quantitative analysis showed that 137 proteins significantly differentially expressed during early maize grain development, and these proteins were involved in protein metabolism, stress response, cell growth and division, carbohydrate and energy metabolism, transport, secondary metabolism, starch synthesis, transcription, lipid metabolism, signal transduction and amino acid metabolism. Among them, the proteins expressed more differentially were related to protein metabolism, stress response, cell growth and division, carbohydrate and energy metabolism. Expression pattern clustering analysis showed that these proteins in different functional categories expressed synergically to regulate the early development of maize grain.

Selection and Cloning of Thaumatin-like Protein (TLP) Gene from Winter Brassica rapa and Its Expression under Low Temperature Stress

MA Li,YUAN Jin-Hai,SUN Wan-Cang,LIU Zi-Gang,ZENG Xiu-Cun,WU Jun-Yan,FANG Yan1,LI Xue-Cai,CHEN Qi,XU Yao-Zhao,PU Yuan-Yuan,LIU Hai-Qing,YANG Gang,LIU Lin-Bo

Acta Agron Sin. 2017, 43(04):  620-628.  doi:10.3724/SP.J.1006.2017.00620

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Thaumatin-like protein (TLP) is an essential protein induced by low temperature stress and enhances plant resistance to adverse circumstances. In our study, we selected differential protein spots from Longyou 7 leaves under low temperature stress and separated TLP closely related to cold resistance by using two-dimensional electrophoresis and mass spectrometry. Using the primers of TLP cDNA sequences and reverse transcription PCR (RT-PCR), we cloned the complete open reading frame of TLP gene of winter Brassica rapa cultivar Longyou7. The sequence length of TLP from Brassica rapa was 732 bp, encoding a protein of 243 amino acid residues with a predicted molecular weight of 26.02 kD and a theoretical pI of 9.02. Bioinformatics analysis showed a similarity of 99.18% in TLP amino acid sequence between B. rapa and Brassica napus, the predicted TLP contained a conserved amino acid sequence belonging to the plant GH69-TLP-SF superfamily. The TLP had one signal peptide which belongs to hydrophilic protein. Moreover, the detection of subcellular localization suggested the synthetic process of this protein is in the endoplasmic reticulum. The prediction of the second structures indicated that TLP is a labile protein consisting of random coils and extended strands. The expression analysis of TLP gene by utilizing real time and Semi-quantitative RT-PCR suggested that the TLP expression could be up-regulated in response to lower temperature. In conclusion, TLP gene of the winter Brassica rapa cultivar Longyou 7 might play a significant role in response to cold stress.