Scientia Agricultura Sinica

Expression of Pathogenesis-Related Proteins in the Interactions Between Rice and Xanthomonas oryzae pv. oryzae

GUAN Ming-Li, DOU Shi-Juan, LI Xue-Jiao, JIA Lin, SHI Jia-Nan, ZENG Xiang-Ran, JIA Meng, GUO Mei-Cen, LIU Li-Juan, LI Li-Yun, LIU Guo-Zhen

Scientia Agricultura Sinica. 2013, 46(20): 4179-4188. doi:10.3864/j.issn.0578-1752.2013.20.001

Abstract ( 391 )

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【Objective】The expression profiling and functional characterization of pathogenesis-related (PR) protein will be helpful to understand the mechanism of rice disease resistance.【Method】To better understand the function of PR proteins, ten differentially transcripted PR genes identified in our previous work using whole genome rice microarray were selected. Western blotting (WB) analysis were used to survey the expression profile in normal growth leave and rice-Xanthomonas oryzae pv. oryzae (Xoo) interactions.【Result】The results indicated that the expression of Os12g43380, Os12g36830, Os12g36860, Os12g36840, Os02g41670, Os05g35290 and Os12g33610 increased gradually during leave growth, reached the highest peak at flowering stage and decreased slightly at mature stage. In rice-Xoo interactions, the expression of Os01g51570, Os01g71680 and Os12g36850 were up-regulated and the expression of Os12g36830, Os12g36840, Os02g41670 and Os05g35290 were down-regulated. It is interesting to note that the expression pattern of PR proteins was similar in both compatible and incompatible rice-Xoo interactions. In addition, cis-elements analysis of the promoter region of PR genes revealed possible defense-related cis-elements, ARE, HSE, MBS and TC-rich repeats were found more frequently in the upstream region of defense-related PR genes.【Conclusion】Seven differentially expressed PR proteins in rice-Xoo interactions were identified.

Advances in Study of Quality Property Improvement of Winter Wheat in China

WEI Yi-Min-1, ZHANG Bo-1, GUAN 二Qi-1, ZHANG Guo-Quan-2, ZHANG Ying-Quan-1, SONG Zhe-Min-3

Scientia Agricultura Sinica. 2013, 46(20): 4189-4196. doi:10.3864/j.issn.0578-1752.2013.20.002

Abstract ( 366 )

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Wheat, being a third grain crop in China, plays a significant role in agricultural and food industries. In China today, economic and social development have greatly affected the living standard of consumers due to industrialization and urbanization of the food industry，especially the traditional food industry, which has developed rapidly. In addition, the food industry requires specialization and large scale (variety and quality) production of food products with a high standard level. Studying and proffering solution to this problem pose a challenge for the sustainable development of wheat production and food industry. This paper has reviewed the existing literatures of home in order to analyze the history, research advantages and recent problems of wheat quality breeding. The requirements of wheat quality for sustainable development of wheat production chain and food industry are discussed. The recommendations proposed in this paper will be useful for departments of wheat breeding，production, storage and food industry as well as agricultural scientists.

Genetic Diversity Research of Mungbean Germplasm Resources by SSR Markers in China

LIU Yan, CHENG Xu-Zhen, WANG Li-Xia, WANG Su-Hua, BAI Peng, WU Chuan-Shu

Scientia Agricultura Sinica. 2013, 46(20): 4197-4209. doi:10.3864/j.issn.0578-1752.2013.20.003

Abstract ( 473 )

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【Objective】The purpose of this research was to investigate the genetic diversity, relationship and differentiation of cultivated mungbean germplasm resources in China, and provide useful information on effective utilization of resources, new genes mining and breeding.【Method】The genetic diversity of 18 mungbean groups from different areas was analyzed by using 40 SSR markers.【Result】A total of 125 alleles were identified with an average of 3.1 alleles per locus. The average of effective number of alleles (NE), Nei's gene diversity (H), polymorphism information content(PIC), expected heterozygosity (He), and Shannon’s information index (I) were 1.8, 0.4233, 0.3497, 0.4241, and 0.6754, respectively. Hebei, Shandong and Anhui provinces had a higher genetic variation than other provinces. The average observed heterozygosity (Ho) was 0.1001 and the average of inbreeding coefficient among population(Fis) was 0.6759, indicating that a certain degree of inbreeding existed in Chinese mungbean germplasm. The gene flow (Nm= 0.6936) and the average genetic differentiation (Fst= 0.2649) also showed a high genetic variation within Chinese mungbean. A total of 272 individuals were divided into 2 groups and 18 populations were divided into 3 groups based on Popgene software. The genetic distances between groups were agreeable with their geographical origin.【Conclusion】There was a high genetic diversity level within Chinese mungbean germplasm. Geographical environment and ecological factors affect the genetic differentiation among different populations.

Breeding Introgression Potato Lines with Resistance to Cold-Induced Sweetening and Screening for Chip Processing Lines

ZHAO Qing-Xia-12, LIN Bi-Bo-12, ZHANG Xin-12, LI Hui-Jun-12, LIU Yue-Shan-2, XU Gang-12, CHENG Li-Xiang-12, WANG Yu-Ping-2, ZHANG Jun-Lian-2, WANG Di-2, ZHANG Feng-12

Scientia Agricultura Sinica. 2013, 46(20): 4210-4221. doi:10.3864/j.issn.0578-1752.2013.20.004

Abstract ( 329 )

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【Objective】The potato introgression lines were bred from hybridization and continuous backcrossing between the primitive and wild varieties with 2n gametes species and the main local cultivated potato varieties, so as to select the chip processing lines and the best introgression parents. 【Method】 The progenies were screened by their field agronomic traits three years at different ecological areas. The content of reducing sugars, free amino acids, acrylamide and activity of acid invertase were analyzed after storage at room and low temperatures and correlation analysis among chip colors and these data were conducted. 【Result】The introgression lines from S. phureja and S. chacoense had higher yield and dry matter content. The introgression lines showed the significantly different ability to accumulate reducing sugars through low temperature storage (4℃). Introgression lines 0712-33, 0722-90, 0732-43 and 0742-66 had low levels of reducing sugars, acrylamide content and acceptable chipping colors. 【Conclusion】The transfer of primitive and wild species’ processing traits into local cultivars by hybridization and continuous backcrossing is an effective potato breeding method and that the germplasm resources S. phureja and S. chacoense are suitable for improving the processing traits of local varieties.

Effects of Seeding Date and Variety Type on Yield, Growth Stage and Utilization of Temperature and Sunshine in Rice

XU Ke-1, SUN Zhen-12, HUO Zhong-Yang-1, DAI Qi-Gen-1, ZHANG Hong-Cheng-1, LIU Jun-2, SONG Yun-Sheng-1, YANG Da-Liu-1, WEI Hai-Yan-1, WU Ai-Guo-2, WANG Xian-2, WU Dong-Dong-1

Scientia Agricultura Sinica. 2013, 46(20): 4222-4233. doi:10.3864/j.issn.0578-1752.2013.20.005

Abstract ( 472 )

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【Objective】This study was conducted to investigate the effects of seeding date and variety type on yield, growth stage and utilization of temperature and sunshine under the rice-wheat cropping system in midland of Jiangsu Province. The result will provide a theoretical basis for scientific use of the seeding date and adoption of suitable variety types.【Method】With medium-maturing late Japonica rice varieties, early-maturing late Japonica rice varieties, late-maturing medium Japonica rice varieties, medium-maturing medium Japonica rice varieties and late-maturing medium Indica rice varieties as materials, using four seeding dates, a comparative study of seeding date and variety type on yield, growth stage and utilization of temperature and illumination was conducted.【Result】The grian productivity of different variety types showed that late-maturing medium Japonica rice varieties＞early-maturing late Japonica rice varieties＞medium-maturing medium Japonica rice varieties＞late-maturing medium Indica rice varieties＞medium-maturing late Japonica rice varieties. With delay of seeding date, the grain yields of the five rice variety types all decreased significantly, but the decrements varied. The yields decreasing range of different variety types showed that medium-maturing late japonica rice varieties＞late-maturing medium indica rice varieties＞early-maturing late japonica rice varieties＞late-maturing medium japonica rice varieties＞medium-maturing medium japonica rice varieties. With the delay of seeding date, the growth process of all the varieties delayed and the growth stages of all the variety types shortened, the accumulated temperature and illumination hours for growth stages significantly reduced. In the same seeding date, the yield discrepancy among different varieties showed an increasing trend with the delay of seeding date. According to the growth stage, the effect of seeding date on the utilization rate of temperature and light resources of different variety types could be divided into two categories, one was the little influence on the short growth period which were medium-maturing medium japonica rice varieties and late-maturing medium indica rice varieties，the other was the great influence on the long growth period which were late-maturing medium japonica rice varieties, early-maturing late japonica rice varieties and medium-maturing late japonica rice varieties. The suitable variety types were preliminarily regionalized based on the different seeding dates as follows: early-maturing late japonica rice varieties and late-maturing medium japonica rice varieties, medium-maturing medium japonica rice varieties, late-maturing medium indica rice varieties should be adopted in Mid-May; late-maturing medium japonica rice varieties and early-maturing late japonica rice varieties, medium-maturing medium japonica rice varieties, late-maturing medium indica rice varieties should be adopted in late May; late-maturing medium japonica rice varieties and medium-maturing medium japonica rice varieties and early-maturing late japonica rice varieties should be adopted in early June; medium-maturing medium japonica rice varieties should be adopted in mid-June. 【Conclusion】 With the delay of seeding date, the grain yield, days of growth stage and utilization of temperature and illumination of the five rice variety types all decreased. Considering the rice-wheat cropping systems and heat conditions, a preliminary division for adopting suitable variety types of different seeding date was made, in order to provide reference for rice production in large area.

The Temporal Variation Characteristics and Spatial Distribution Laws of Drought of Spring Maize in Northern China

DONG Chao-Yang-1, YANG Xiao-Guang-1, YANG Jie-1, JIE Wen-Juan-12, YE Qing-13, ZHAO Jin-1, LI Ke-南1

Scientia Agricultura Sinica. 2013, 46(20): 4234-4245. doi:10.3864/j.issn.0578-1752.2013.20.006

Abstract ( 398 )

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【Objective】 The tendency of inter-decadal evolution characteristics and spatial distribution laws of drought of spring maize at each growth stage in northern China were studied. 【Method】 Based on the daily meteorological data and spring maize growing records of 291 meteorological stations during 1961-2010 in the study area, the tendency of inter-decadal evolution characteristics and spatial distribution laws of drought of spring maize in the study area were analyzed using the crop water deficit index as the agricultural drought index.【Result】As for the CWDI of spring maize, the fluctuations in different years in Northwest China was steady, while in the other two areas in 1980s and 1990s the fluctuations were relatively severe. And the CWDI of spring maize at the stage of tasselling to maturation in North China was lower than the other two stages significantly, while the changes in Northeast China and Northwest China at different growth stages were not obvious. There was a significant fluctuation of the classification of drought during different decades in the central region of North China. The spatical distributions of the classification and frequency of drought of spring maize in northern China showed higher in west and north and lower in east and south, they both had the highest value in Northwest China, followed by North China, and the lowest value in Northeast China. Results showed a significant spatial distribution of the frequency of extreme drought and light drought. The extreme drought was mainly concentrated in most of Xinjiang, the north part of Gansu and the northwest reigon of Inner Mongolia. It occurred two times or above in three years. While the light drought was mainly concentrated in most of Northeast China, most of North China and the southeast part of Northwest China. It occurred one time or above in five years. Moderate drought and severe drought were mainly concentrated in North China and the east part of Northwest China. They occurred one time or above in five years, and they had a decreasing trend with growth stages changing. 【Conclusion】 As for the CWDI of spring maize in northern China, the fluctuations in different years in North China and Northeast China were relatively severe, and there was an upward trend since 1980s. The spatical distributions of the classification and frequency of drought indicated a significant east-west distribution. The extreme drought happened in higher frequency in the west and lower in the east, and it has expended in the stage of tasselling to maturation, on the contrary, the light drought occurred in higher frequency in the east and lower in the west, and it had increased at the stages of tasselling to maturation. As for the moderate drought and severe drought, they both showed a higher frequency in central regions and a lower frequency in east and west regions, and they decreased both in area and degree at the stages of tasselling to maturation. The sensitive area of drought grade variation at different growth stages were the north part of Xinjiang and the central region of North China.

Effect of Phosphatidylinositol 3-Phosphate on UV-B-Induced H2O2 Production in Guard Cells and Stomatal Closure of Broad Bean

LI Hui-Min-12, HU Jie-1, HE Jun-Min-1

Scientia Agricultura Sinica. 2013, 46(20): 4246-4253. doi:10.3864/j.issn.0578-1752.2013.20.007

Abstract ( 371 )

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【Objective】 The role of phosphatidylinositol 3-phosphate (PI3P), the product of phosphatidylinositol 3-kinase (PI3K), in UV-B-induced hydrogen peroxide (H2O2) generation in guard cells and stomatal closure was analyzed to provide valuable evidence for further elucidating the mechanism of UV-B signaling transduction in plants. 【Method】 By using PI3K inhibitors wortmannin (WM) and LY294002 (LY) to inhibit the production of PI3P, and by using diphenylene iodonium (DPI) and salicylhydroxamic acid (SHAM) to inhibit H2O2 production via NADPH oxidase pathway and peroxidase pathway respectively, the role of PI3P in 0.8 W?m-2 UV-B-induced H2O2 production in guard cells and subsequent stomatal closure was studied via stomatal bioassay and laser-scanning confocal microscopy in epidermal strips of broad bean (Vicia faba L.). 【Result】Both the UV-B-induced H2O2 production in guard cells and stomatal closure were significantly inhibited by WM and LY. Exogenous H2O2 rescued the inhibitory effect of WM and LY on UV-B-induced stomatal closure, but exogenous H2O2-induced stomatal closure was not inhibited by WM and LY. Both the UV-B-induced H2O2 production in guard cells and stomatal closure were significantly inhibited by the reactive oxygen species scavenger and SHAM, but not inhibited by DPI. 【Conclusion】 PI3P mediates UV-B-induced stomatal closure via activating the peroxidase-dependent H2O2 production in guard cells of broad bean.

Integrated Management of Wheat Stripe Rust Caused by Puccinia striiformis f. sp. tritici in China

CHEN Wan-Quan-1, KANG Zhen-Sheng-2, MA Zhan-Hong-3, XU Shi-Chang-1, JIN She-Lin-4, JIANG Yu-Ying-5

Scientia Agricultura Sinica. 2013, 46(20): 4254-4262. doi:10.3864/j.issn.0578-1752.2013.20.008

Abstract ( 773 )

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Wheat stripe (yellow) rust caused by Puccinia striiformis f. sp. tritici (Pst) is an important disaster in wheat production. A great deal of effort has been spent on the strategies and techniques of integrated management of wheat stripe rust for over six decades by a teamwork nationwide in China. It has been fully revealed that the over-summering and over-wintering areas of Pst inter-regional dispersal of inoculum, the way of virulence variation, and breakdown causation of wheat cultivar resistance. Based on the geographical conditions, wheat growth, occurrence and dispersal of disease etc, wheat stripe rust in China can be divided into three major zones, namely the autumn sources of inocula, the spring sources of inocula, and spring epidemic areas. A major strategy of headstream management has been put forward, i.e. ‘integrated management of wheat stripe rust in the sources of inocula to protect wheat safe plantation in the whole country. A series of effective measures for the forecast and control of disease has been developed, which include the molecular diagnosis of stripe rust, remote wide forecast, improving cultivar resistance, changing cultivation crops, regulating sowing date, seed-dressing with fungicides, and spraying fungicides at the initial stage of disease, etc. According to the epidemic rules of wheat stripe rust, the integrated management systems based on the biodiversity have been set up in the areas of inoculum sources of Pst, respectively, which have been widely applied in wheat production resulting in the sustainable control of wheat stripe rust epidemics and remarkably economic efficiency. In this paper, it was also proposed as the priority that variation mechanism of pathogen virulence, development of early forecast system and the ecological control measures of disease in the areas of inoculum sources of wheat stripe rust in the near future.

Cloning and Localization of an Odorant Receptor Gene OR18 in the Antenna of Spodoptera exigua

LIU Cheng-Cheng-12, LIU Yang-2, ZHANG Jin-12, WANG Gui-Rong-2, DONG Shuang-Lin-1

Scientia Agricultura Sinica. 2013, 46(20): 4263-4271. doi:10.3864/j.issn.0578-1752.2013.20.009

Abstract ( 437 )

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【Objective】 The objective of this study is to clone odorant receptor gene and investigate its expression in different tissues and antennal sensilla of Spodoptera exigua adults, and thus to explore the function of the odorant receptor gene. 【Method】 The full length cDNA sequence encoding SexiOR18 was cloned by homologous-based PCR and RACE techniques. The expression level of this gene in different tissues was investigated by quantitative real-time PCR. The in situ hybridization was used to examine its localization in different antennal sensilla of adult antennae.【Result】 A 740 bp fragment of a OR candidate gene was obtained from the antenna of S. exigua by using homologous-based cloning strategy. The full-length cDNA sequence encoding this gene was further obtained by using RACE technique. The gene was named as SexiOR18 (GenBank accession number: JN873314). The full-length cDNA of SexiOR18 is 1 618 bp with an open reading frame of 1 194 bp, which encodes a polypeptide of 398 amino acids. The multiple alignment and phylogenetic tree analyses showed that SexiOR18 shared high identity with the homologs from other Lepidopteran species, especially noctuid insects. The expression measurements revealed that SexiOR18 expressed in significantly higher level in female antenna than in male antenna, and within the antenna, the expression was mainly restricted under trichoid sensilla and basiconic sensilla, but no expression under coeloconic sensilla and chaetica sensilla.【Conclusion】The SexiOR18 localizes under both trichoid sensilla and basiconic sensilla, suggesting its involvement in detection of both pheromone and general odorants.

Cloning and Functional Analysis of pdp1 in Ostrinia furnacalis (Lepidoptera: Crambidae)

CHENG Xiao-Juan-12, YAN Shan-Chun-1, HUANG Yong-Ping-2, TAN An-Jiang-2

Scientia Agricultura Sinica. 2013, 46(20): 4272-4283. doi:10.3864/j.issn.0578-1752.2013.20.010

Abstract ( 339 )

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【Objective】The objectives of this study are to clone pdp1 (protease-activated receptor-domain protein1 gene) which named Ofpdp1 (GenBank accession number: KC857457) from the Asian corn borer, Ostrinia furnacalis (Guenée), and to identify its typical hallmarks, then to analyze the mRNA expression characteristics and physiological function of Ofpdp1. It will provide potential target gene for genetic control of O. furnacalis.【Method】 The cDNA sequence of Ofpdp1 was isolated using RT-PCR and PCR methods. Based on the sequencing results, the bioinformatics analysis of nucleic acid and putative amino acid was conducted. The developmental expression pattern of Ofpdp1 was determined by qPCR. RNAi experiment was performed at proembryo stage to explore the physiological function. 【Result】The cDNA sequence (960 bp) of OfPDP1 was obtained, including a complete open reading frame (ORF) of 804 bp, which encoded 267 amino acids. OfPDP1 was a transcription factor of PAR bZIP (basic leucine zipper) subfamily which included several conserved regions in the carboxy terminal region of the protein. The result of phylogenetic analysis was consistent with the classification of the species in biology. The qPCR results showed a high level expression at the 76 h of eggs’ development process. It could be declined by 71% in the transcript levels of Ofpdp1 in the eggs injected with Ofpdp1-specific dsRNA in 48 h compared to those in the control eggs. 【Conclusion】 A complete ORF of Ofpdp1 was sequenced with clone strategy from Asian corn borer for the first time. Ofpdp1 was highly conserved in the process of evolution, and it belonged to PAR bZIP subfamily. The high levels expression of Ofpdp1 at the time of 72 h suggested that Ofpdp1could take an important part in the changing process of embryo physiological structure during the larva’s forming period.

Influence of Sprayer and Application Rate on Pesticide Deposit Character on Rice Canopy

XU De-Jin, GU Zhong-Yan, XU Guang-Chun, XU Xiao-Long

Scientia Agricultura Sinica. 2013, 46(20): 4284-4292. doi:10.3864/j.issn.0578-1752.2013.20.011

Abstract ( 370 )

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【Objective】The objective of this study is to illustrate the relationship between pesticide deposit structure and spatial distribution on control efficiency, and to study the effects of sprayer and application rate on pesticide deposit character on rice canopy. 【Method】Droplets coverage and density of pesticide were assayed with DepositScan software after pesticide collected with the relevant collection device and water-sensitive paper. Meanwhile, the tracer material was also used to estimate the amounts of pesticide deposition.【Result】Sprayed by the motorized knapsack mist blower (MKM) and the lever-operated knapsack sprayer (LKS) at different application rates, the droplet coverage rate of pesticide at adaxial side of rice leaf was significantly higher than that at abaxial side with a leaf angle of 45° and 0°. The droplet densities at the abaxial side of the leaf with a leaf angle of 0° were more than 200 droplets/cm2 with pesticide sprayed by the MKM at the application rates of 225, 450 and 600 L•hm-2. Moreover, significant differences were found among these application rates. The droplet densities at the abaxial side of the leaf with the leaf angles of 0 ° and 45° were all below 15 droplets/cm2 with pesticides sprayed by LKS at the application rates of 600, 900 and 1 200 L•hm-2. In addition, no significant differences were found among these application rates. Application with the same dosage, the pesticide deposit amount in rice canopy at different sites reached the summit with pesticides sprayed by LKS at the application rate of 450 L•hm-2. However, a significant difference was not observed among the pesticide deposit amounts with pesticides sprayed by LKS at different application rates. Compared with MKM, pesticide deposit amounts at the leaf abaxial side were lower with pesticides sprayed by LKS. Application of chlorantraniliprole at 15 g active ingredients per hm2, the best control efficiency was observed at the application rate of 450 L•hm-2 sprayed by MKM, however, the worst control efficiency was showed at the application rate of 1 200 L•hm-2 sprayed by LKS.【Conclusion】 Application with both MKM and LKS in rice field, the pesticide droplets coverage rates at leaf adaxial side were all higher than leaf abaxial side. Sprayed by MKM, improvements of the droplets density and coverage rate were observed at the leaf abaxial side with the increase of the pesticide application amounts. However, sprayed by LKS, significant effects were not found in this study. Compared with LKS, solution sprayed by MKM can increase the droplet density, coverage rate and the pesticide deposit amount significantly, thus enhancing the control effect.

Establishment and Validation of Model of Soil Particle Size Distribution of Main Soils in China by Laser Diffraction Method

WU Huan-Huan-12, 吕Jia-Long-2 , DUAN Ying-Hua-1, ZHANG Wen-Ju-1, XU Ming-Gang-1

Scientia Agricultura Sinica. 2013, 46(20): 4293-4300. doi:10.3864/j.issn.0578-1752.2013.20.012

Abstract ( 380 )

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【Objective】 To improve the accuracy of method measuring soil particle size distribution, the new method was studied. 【Method】 Twenty soil samples collected from 15 provinces were determined by Laser Diffraction Method and Pipette Method. 【Result】 Results show that a significant relationship was found between the clay sand contents determined by two different methods. The clay and sand content determined by using the Laser Diffraction Method were obviously lower than that by using the Pipette Method (73.4 g•kg-1 and 59.6 g•kg-1), while the silt content by using the former method was higher than that by using the latter (131.2 g•kg-1). A new alternative model between the two methods was found by studying the particle size cumulative distribution. If the function fitted by particle size cumulative distribution by using Laser Method of a soil is y=1 000x/(b+x), then the function by using Pipette Method is y=1000x/(1.32b-1.01+x). 【Conclusion】 The soil texture can be judged by the value calculated based on the latter function. The findings demonstrate that data from the Laser Diffraction Method can be directly used to classify soil texture.

Construction of the Suppression Subtractive Hybridization Library and Analysis of Related Genes of Spur-Type Apple (Malus domestica Borkh.)

SONG Yang-12, WU Shu-Jing-1, ZHANG Yan-Min-1, CHEN Xue-Sen-1

Scientia Agricultura Sinica. 2013, 46(20): 4301-4309. doi:10.3864/j.issn.0578-1752.2013.20.013

Abstract ( 347 )

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【Objective】The aim of this experiment is to explore genes related to spur type shoots of apple and to try to understand the mechanism of sport for breeding new spur type variety.【Method】A forward and reverse suppression subtractive hybridization library were constructed using the spur type shoots from sport and the standard shoots. Identification of the inserted cDNA fragments in subtracted library was done using real-time PCR.【Result】A total of 291 positive clones were obtained and the size of the inserts was 300 to 600 bp. By means of BLAST comparison of GenBank, 126 ESTs were found to share considerable homology with known genes while the rest 165 ESTs had no homology with known genes. The putative function of these genes was involved in metabolism, transcription, stress response, transport, photosynthesis, and signal transduction. The ESTs were significantly differently expressed between the spur type sport and the standard shoots. 【Conclusion】 Some genes related to shoot internode length of spur type apple were obtained through construction of the forward and reverse suppression subtractive hybridization library. The research results suggested that the genes play an important role during the shoots elongation of spur type apple.

Changes of Qualities During the Fruit Late Development of ‘Golden Delicious’ Apple and Distinguishing Its Ripening Process

WANG Hai-Bo, LI Lin-Guang, LIU Jia-Fen, HE Ping, LI Hui-Feng, ZHANG Yong, YANG Jian-Ming

Scientia Agricultura Sinica. 2013, 46(20): 4310-4320. doi:10.3864/j.issn.0578-1752.2013.20.014

Abstract ( 452 )

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【Objective】 Critical time of fruit quality formation and distinguishing of ripening process of ‘Golden Delicious’ apple were studied, aiming to offer a theoretical basis for determining its proper harvest time and realizing its high-quality production.【Method】Basic fruit quality parameters, including fruit weight, total soluble solids (TSS), soluble sugars, titratable acid, sugar-acid ratio, vitamin C (Vc), flesh firmness were investigated using conventional methods. Volatile compounds were identified by static headspace and gas chromatography-mass spectrometry. Sugars and organic acids were quantified using high-performance liquid chromatography. 【Result】 During the late development of the cultivar, fruit weight, TSS, total sugars, sugar-acid ratio, volatile eaters and sugar constitutes increased, while titratable acid, Vc, flesh firmness and acid constitutes except malic acid decreased, and volatile alcohols, aldehydes and malic acid increased first and decreased afterwards, generally. Basic quality parameters changed steadily 168 days after full bloom (AFB) in disregard of the faster increase of TSS and decrase of flesh firmness from 182 to 189 d AFB. From 161 d AFB, acetate and butyrate esters began increase and C6 aldehydes began decrease rapidly. Alcohols increased rapidly from 161 to 168 d AFB and decreased rapidly after 168 d AFB. Sugar constitutes increased steadily after 168 d AFB. 161 d AFB was the time of malic acid from rising to falling. The results of principal component analysis of all the variables in this work indicated the stage change of tested parameters and ripening process.【Conclusion】Major change period of basic quality parameters and sugar constitutes was before 168dAFB, and major change period of esters, aldehydes, alcohols and malic acid was after 161d AFB. The period of late development of ‘Golden Delicious’ could be distinguished three stages: unripe (140-147 d AFB), early ripe (154-161 d AFB), and ripe (168-189 d AFB).

Effects of Phenanthrene and Pyrene on Ca2+, K+ and H+ Liquidity in Root Hair Cells of Malus hupehensis

HUANG Cui-Xiang, XIA Yan-Fei, WANG Rong, ZHANG Wen-Hui, HAN Tian-Tian, SHEN Xiang

Scientia Agricultura Sinica. 2013, 46(20): 4321-4327. doi:10.3864/j.issn.0578-1752.2013.20.015

Abstract ( 314 )

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【Objective】 The objective of this study is to prove PAHs to the plant growth mechanism, and the difference in substances poisoning effect of different PAHs. 【Method】 The Non-invasive Micro-test Technique was used to measure in Malus hupehensi sweet tea seedling root treated by using the polycyclic aromatic hydrocarbon phenanthrene and pyrene treated Ca2 +, K +and H +velocity variation. 【Result】 The Ca2 +, K + and H + liquidity dramatically reversed by contrasting from (-63.53±9.30） pmol•cm-2•s-1, （-60.56±14.56） pmol•cm-2•s-1, （+44.38±5.19 ）pmo•cm-2•s-1, changing to (+127.18±39.95) pmol•cm-2•s-1, (109.97±25.68) pmol•cm-2•s-1, and (-10.35±1.57 ) pmol•cm-2•s-1) treated with phenanthrene; and treated with pyrene, the Ca2+ , K +and H + liquidity was reversed by changing to （+220.29±60.42） pmo•cm-2•s-1,（140.21±27.87） pmol•cm-2•s-1, and（-14.42±3.16） pmol•cm-2•s-1. 【Conclusion】 PAHs can destroy the cell membrane permeability, reduce the ion channel activity and disturb the Ca2+ and K + ion absorption process of M. hupehensis, thus resulting in the lack of relevant elements, and pyrene can cause poisoning effect which is significantly higher than phenanthrene, and PAHs toxic to plants has become one of the mechanism. The conclusion was that the influence of polycyclic aromatic hydrocarbon on plant growth can be traced back to ion absorption and ion channel change for further study the influence of PAHs on plants and provide a theoretical basis of the influence.

Interspecific Hybridization Between Chrysanthemum morifolium ‘Nannongyinshan’ and C. zawadskii and Identification of Waterlogging Tolerance of Their Hybrid

WANG Liang, WANG Chu-Chu, JIANG Jia-Fu, CHEN Su-Mei, FANG Wei-Min, TENG Nian-Jun, GUAN Zhi-Yong, LIAO Yuan, CHEN Fa-Di

Scientia Agricultura Sinica. 2013, 46(20): 4328-4335. doi:10.3864/j.issn.0578-1752.2013.20.016

Abstract ( 459 )

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【Objective】To improve the waterlogging tolerance of spray cut chrysanthemum, an interspecific hybridization was combined between Chrysanthemum morifolium ‘Nannongyinshan’ and C. zawadskii. The phenotypic characteristics and waterlogging tolerance of the parents and the hybrid were identified. 【Method】Interspecific hybridization was made between the female parent C. morifolium ‘Nannongyinshan’, which is susceptible to waterlogging ,and the male parent C. zawadskii, which is tolerant. The hybridity was identified by morphological traits and SRAP molecular markers. Waterlogging tolerance was investigated by changes in leaf relative water content (RWC), contents of chlorophyll (Chl) and malondialdehyde (MDA), and activities of antioxidant enzymes. 【Result】 Only one putative hybrid was obtained through the interspecific hybridization. Morphological characteristics including the plant height, leaf length, leaf width, number of tubular florets and ray florets of the hybrid were different with that of the parents. In the hybrid, both the male and the female specific bands were observed by SRAP molecular markers analysis. Thus the progeny was a real interspecific hybrid. The decrease of RWC and chl and the increase of MDA of the hybrid were significantly lower than that of maternal ‘Nannongyinshan’, while the activities of antioxidant enzymes were higher. 【Conclusion】 The results indicated that the hybrid was more tolerant to waterlogging than ‘Nannongyinshan’. Therefore, it is possible to obtain new waterlogging tolerant cut chrysanthemum germplasm via interspecific hybridization to introduce the tolerant trait of C. zawadskii.

Relationship Between Wheat Quality and Physicochemical Properties of Extruded Expansion Product

CHEN Feng-Liang, CHEN Xiang-Yan, WANG Xian-Chang, YIN Jing, ZHAO Xiao-Yan

Scientia Agricultura Sinica. 2013, 46(20): 4336-4343. doi:10.3864/j.issn.0578-1752.2013.20.017

Abstract ( 397 )

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【Objective】The relationship between the wheat quality of different varieties and physicochemical properties of corresponding extruded expansion product were investigated to explore which quality properties of wheat is suitable for producing extruded expansion food. 【Method】Wheat expansion products were extruded using lab-scale twin screw extruder from 16 varieties under unified process parameters condition, the quality properties of wheat, the physicochemical properties of expansion extrudate and the relationship between both were analyzed. 【Result】 There were obvious differences in quality properties of wheat and physicochemical properties of corresponding extruded expansion products. There were significant negative relationships between color difference and thousand-kernel weight, flour whiteness, weaken extent, water absorption index and kernel protein content, sedimentation value, wet gluten content, forming time, water solubility index and kernel protein content, wet gluten content, gluten index, stretching length, degree of gelatinization and kernel protein content, sedimentation value, wet gluten content. There were significant positive relationships between color difference and kernel hardness, falling number, water absorption index and volume weight, water solubility index and gluten index, tensile resistance at 5cm, product hardness and kernel hardness, flour whiteness. In the selected 16 wheat varieties, Tainong 18, Jimai 22, Qingfeng 1, Qingnong 2, Yannong 23 and Lumai 21 were more suitable for making extrusion expansion products.【Conclusion】The thousand kernel weight, volume weight, kernel hardness, flour whiteness, kernel protein content, sedimentation value, wet gluten content, gluten index, falling value, forming time, weaken extent, stretching length and tensile resistance at 5 cm all have significant effects on physicochemical properties of extruded expansion product. The wheat varieties or raw materials, with high value in thousand kernel weight, volume weight, gluten index, weaken extent and tensile resistance at 5 cm, low value in kernel hardness, flour whiteness, kernel protein content, sedimentation value, wet gluten content, falling value, forming time and stretching length, were more suitable for producing expansion products.

Comparative Studies on the Bacteriostatic Activity of the Three Bacillus Strains and Its Effects on Air Microorganisms in Broiler House

TONG Yong-Juan-1, SA Ren-Na-1, ZHANG Hong-Fu-1, SUN Chun-Yang-1, XING Huan-1, PENG Yi-Zhu-1, WEI Zhong-Hua-2, WANG Xue-Jing-2

Scientia Agricultura Sinica. 2013, 46(20): 4344-4353. doi:10.3864/j.issn.0578-1752.2013.20.018

Abstract ( 422 )

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【Objective】The present study was conducted to evaluate the biological characteristics of 2 strains of Bacillus amyloliquefaciens and 1 strain of Bacillus thuringiensis, and the bacteriostatic activity of these three bacillus strains against Escherichia coli. After being sprayed the mixture of three bacillus suspension to the broiler house, the number of E.coli and staphylococcus aureus in the air was detected in order to evaluate the effect of bacillus on improvment of environment.【Method】In this study, the colony morphology and the growth curve of the three bacillus were examined. All of the three bacteria were examined for their antimicrobial activity against E. coli (CVCC1570) by using the agar diffusion system. The optimum combination level of the three bacteria in the mixed product was investigated by using the orthogonal design in three factors and three levels. The mixed product of the three bacillus strains was sprayed in the poultry house. Three houses were selected, one for blank control(A), one for chemical disinfectant (benzalkonium chloride) as positive control(B), and one for bacillus treatment(C). Each treatment contained three replications. The numbers of E.coli and Staphylococcus aureus in the air of the poultry house were detected every week.【Result】The results showed that all strains appeared as Gram-positive and aerobic bacillus. Bacillus SKN01, Bacillus SKN02 had the similar growth curve entering the logarithmic phase in the first two hours. Bacillus SKN03 grew slowly and spent long time to enter the logarithmic phase. The effective bacteriostatic activity concentration of single bacillus strain against E. coli (CVCC1570) was 108 CFU•mL-1. The most effective concentration of antimicrobial activity in the mixture of three bacillus strains was 4×108CFU•mL-1, 4×108CFU•mL-1 and 6×108CFU•mL-1. Spraying the mixture of bacillus suspension in the broiler house could effectively reduce the number of E.coli and S. aureus in the air, especially at the later growth stage of chickens.【Conclusion】The bacillus strain had antimicrobial activity against E.coli. Spraying the mixture of the three bacillus suspension in the broiler house could reduce the number of E.coli and S. aureus in the air and could improve the environment effectively.

Comparison of Yields and Nutrient Compositions Between Different Harvesting Heights of Maize Stover

ZHAO Hong-Xiang, NING Tang-Yuan, NIE Liang-Peng, WANG Bing-Wen, TIAN Shen-Zhong, LI Zeng-Jia

Scientia Agricultura Sinica. 2013, 46(20): 4354-4361. doi:10.3864/j.issn.0578-1752.2013.20.019

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【Objective】This study analyzed the nutrients composition, water content and yield of different cutting heights of maize stover in attempt to provide a reference for selection of the best cutting height to feed the ruminants.【Method】A field experiment was carried out in the Huang-Huai-Hai region of a typical wheat-maize cropping system. After maize was harvested, its stover was cut at different heights, including above 100 cm, above 50 cm, above 0 cm, below 100 cm and below 50 cm from the ground. Some physiological indexes of different heights of maize stover were analyzed, respectively, which included moisture content (MC), dry weight (DW), fresh weight (FW), content and yield of crude protein (CP), crude fat (EE), neutral and acid detergent fiber (NDF and ADF), starch (SC), soluble sugar (SS), calcium (Ca), phosphorus (P) and potassium (K) of different treatments, respectively.【Result】With the increase of harvesting height, the content of CP, EE, Ca and P gradually increased, but the contents of MC, SS, NDF, ADF, SC and K gradually reduced, respectively. The production of DW, CP, NDF, ADF, SC and K accounted for about 30% of the total production, respectively; however, the production of Ca, EE and P accounted for about 40% of total production in the above 100 cm treatment, respectively. The production of FW and K accounted for about 50 percent of the total production, the production of DW and nutrients composition except K accounted for 60% to 70% in the above 50 cm treatment.【Conclusion】 From the content and yields of the nutrients, the above 100 cm treatment is best for forage. Therefore, it is recommended that 100 cm from the ground is a suitable height used for forage when maize is harvested in the mechanical harvesting system.

Advances in Study of Molecular Epidemiology of Porcine Epidemic Diarrhea Virus in China

SHI Biao-2, DONG Shi-Juan-1, ZHU Yu-Min-1, YU Rui-Song-1, LI Zhen-1

Scientia Agricultura Sinica. 2013, 46(20): 4362-4369. doi:10.3864/j.issn.0578-1752.2013.20.020

Abstract ( 460 )

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Porcine epidemic diarrhea (PED) caused heavy economic losses to Chinese pig industry in 2010 and 2011. It was postulated that new variants of porcine epidemic diarrhea virus (PEDV) had played a major role because the vaccine made from previous strains failed to provide efficient protection for infected piglets. This review summarized the investigations on molecular epidemics of PEDV in this period of time in China and made a phylogenetic analysis with published complete genomic sequences. The analysis provided further evidence for the existence a novel genotype PEDV, which was not clustered with Chinese 1986 strains and European CV777 strain. The growing genetic difference between prevailing and vaccine strains should be the cause of vaccination failure as well as high death rate of piglets. Complete genomic sequence analysis showed the length difference of their polymerase (1ab) as well as their spike (S) genes. S gene had the highest mutation rate among the subgenomes, especially within a relatively short period of time. The analysis also implied that the new strains could evolve from domestic strains. Severe PED problem was reported early in 2006, therefore the 2010 to 2012 outbreak of PED could be the result of accumulation of pathogen mutations and adaptation, which also suggested, the importance of evolution surveillance of pathogens by national network.

Development of a RT-PCR Method for Differentiation of the Wild-Type PEDVs and the Attenuated PEDVs

WU Yu-Lu-1, CHENG Qun-1, YU Ling-Xue-1, HOU Yi-Xuan-2, WANG Kang-1, LIU Guang-Qing-1, TONG Guang-Zhi-1, ZHOU Yan-Jun-1

Scientia Agricultura Sinica. 2013, 46(20): 4370-4377. doi:10.3864/j.issn.0578-1752.2013.20.021

Abstract ( 370 )

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【Objective】The objective of this study is to establish an effective tool for differentiating diagnosis of the PEDVs in epidemiological investigations by the method of RT-PCR. 【Method】According to the truncation in the ORF3 gene of PEDV reference strains in GenBank, the wild-type PEDV isolates and attenuated isolates, two pairs of primers were designed. The clinical samples from different farms occurred with diarrhea epidemic were tested, and their ORF3 gene was analyzed. Some representative samples were selected and amplified by the primers ORF3-JD1/2. The procedure of the RT-PCR was perfected. A large number of clinical samples were collected to carry out the specificity and sensitivity test.【Result】The sequences of ORF3 gene of 11 PEDV isolates were obtained. No deletion in ORF3 gene of 9 strains was found, and it indicated that they were wild-type PEDVs. And the results showed 95.8%-97.1% nucleotide sequence homology identity between wild isolates and attenuated ones. The RT-PCR was shown to specifically amplify a 300 bp fragment from the wild-type PEDVs or a 250 bp fragment from the attenuated PEDVs. This method showed no cross-amplification between PEDVs and other porcine virus. And the sensitivity of detection of viral was upto 100 TCID50. The clinical samples were tested by using this RT-PCR method , and the results showed a 65.4% PEDV positive rate.【Conclusion】 The RT-PCR could be used as an effective tool for differentiating diagnosis of the PEDVs in epidemiological investigations.

Effect of Heat Stress on the Intestinal Flora Structure and Alkaline Phosphatase Activities and mRNA Expression of Amino Acid Transporters of Layer

LI Yong-Zhu-1, CHEN Chang-Xiu-1, YongquanCui2

Scientia Agricultura Sinica. 2013, 46(20): 4378-4387. doi:10.3864/j.issn.0578-1752.2013.20.022

Abstract ( 452 )

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【Objective】 The objective of this study is to reveal the influence mechanisms of heat stress affecting the intestinal flora structure of layer, the alkaline phosphatase activities of intestinal mucosa and the mRNA expression of amino acid transporters.【Method】A total of 96 Jining Bairi chicken aged at 16 weeks were chosen and randomly divided into control group (group I, (24±1)℃) and heat stress group ((38±1)℃). six replicates were desighed for each group with 8 individuals for each replicate, and the duration sustained for 14 days. PCR-DGGE and RT-PCR of 16S rDNA were used. The bacterial diversity of the contents in dodecadactylon, jejunum and ileum, alkaline phosphatase activities of intestinal mucosa and the mRNA expression of amino acid transporters e.g. rBAT, y+LAT1, and CATl were analyzed when the layers were under the heat stress on 2nd day (group II), 7th day (group III) and 14th day(group IV), respectively.【Result】The bacterial diversities of each intestinal segments showed relatively abundant from the 7th day after heat stress. On the 7th and 14th days, no sensitive lactobacillus in jejunum and ileum was detected, and so were the L. johnsonii, nonculturable bacteria and Bacteroides in ileum. While the nonculturable bacteria, ulcer Bacteroides, ulcer Bacteroides, and Shigella sonnei were detected at every phase of heat stress. The alkaline phosphatase activities of mucosal epithelial cells in jejunum and ileum decreased significantly compared to the control group (P＜0.05). The mRNA expressions of rBAT, y+LAT1 of the group III in jejunum and ileum were the lowest. The variation extent of the mRNA expression in jejunum at each phase of heat stress was the greatest (P＜0.05). The mRNA expressions of CAT1 in ileum of groups III and IV were affected more greatly by the heat stress compared to that of group I (P＜0.01).【Conclusion】The influence on the intestinal flora in jejunum and ileum by heat stress is more evidently and the intestinal function of digestion and absorption can be changed by the variation of the intestinal flora..

Expression Features of BnMAPK1 in Wound and Pathogenetic Fungi Stress

LU Jun-Xing, LU Qi-Feng, ZHANG Kai, CHAI You-Rong, LI Jia-Na, QIAN Wei, 吕Jun , LU Kun, LIANG Ying

Scientia Agricultura Sinica. 2013, 46(20): 4388-4396. doi:10.3864/j.issn.0578-1752.2013.20.023

Abstract ( 389 )

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【Objective】The objective of this study is to analyze tissue specific expression of BnMAPK1 genes, and explore their characteristics and signaling pathway in wound and pathogenetic fungi stress.【Method】On the basis of silico cloning, the promoters of BnMAPK1 were cloned. BnMAPK1 mRNA expression characteristics after induced were analyzed by Quantitative real-time PCR (qRT-PCR). 【Result】The cloned promoter sequences of BnMAPK1 contain several hormone and stress responsive elements. BnMAPK1 expression could be induced by phytohormones methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA), hydrogen peroxide (H2O2), wound, and Sclerotinia sclerotiorum. Further experiments showed that PDF1.2 gene was activated but PR-1 was suppressed after wound and S.sclerotinrum treatments, indicate that oilseed rape most likely through jasmonic acid (JA) signaling in response to wound and pathogenetic fungi stress.【Conclusion】BnMAPK1 expression could be induced by several phytohormones and its promoter contains several hormone and stress responsive elements. BnMAPK1 may play an important role in resistance to a variety of stress in plants and oilseed rape might be mediated by JA signaling pathway in response to wound and pathogenetic fungi stress.

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