Gene Cloning and Tissue-Specific Microplitis mediator （Hymenoptera： Expression of G Protein β Subunit in Braconidae）

A gene encoding a novel G protein β subunit of β1 subclass, GβMmed was isolated from Microplitis mediator （Hymenoptera： Braconidae）. The full-length sequence of GβMmed is 1 119 bp, the cDNA contains a 1 023 bp open reading frame that encodes a protein with 340 amino acids, and the predicted molecular weight of GβMmed is 37.23 kDa and isoelectric point is 5.86. By the quantitative real-time RT-PCR method, the tissue-specific expression and quantitative changes in the developmental expression profile of GβMmed were detected. It was found that GβMmed was abundantly expressed in M. mediator antennae, head （without antennae）, thorax, abdomen, legs and the wings, and especially at high levels in abdomen. In antennae, expression varied through 1st day before emergence to 5-d-old adults, and had equal expression levels detected in females and males in total. In head, GβMmed expresses while initially high in females, and have another peaked in stage 4 and 1st day, in males showed a peak of GβMmed expression prior to emergence and relatively low levels after emergence. In female abdomen GβMmed expression levels have two peaks in stage 1 and the 5th d, but just have one peak in male abdomen in stage 1. In all other tissues expression was low and stable.     

Cloning of Proteinase Inhibitor Gene StPl in Diploid Potato and Its Expression Analysis

A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum （Rs） resistant potato leaves using the rapid amplification of cDNA ends （RACE） method and designated as StPI. BLAST search against NCBI showed that the StPI gene shared 89% identity with potato proteinase inhibitor I precursor in nucleotide and 74% in amino acid. Analysis of semi-quantitative RT-PCR indicated that this gene was induced by Rs as well as up-regulated by jasmonic acid （JA）. The StPI gene expression reached the highest level during 6-12 h post Rs-inoculation or JA-treatment, and then leveled off. Moreover, this gene was strongly induced by JA and its mRNA accumulation increased more quickly than that of Rs-inoculation. The StPI gene may play a role in potato resistance against Rs. The induction of StPI by Rs invasion may have a similar signal transduction pathway with JA treatment.     

Cloning of Soybean 24 kDa Oleosin Gene and Its Transient Expression asa Carrier for Foreign Protein

The genomic DNA sequence encoding soybean 24 kDa oleosin and its promoter were cloned andanalyzed for investigation of the potentials of the oleosin acted as a carrier for production of recombinant proteins in plant. The -300 box, GA-rich, G-box, SEF-3, SEF-4, RY box, ABA box, CAn and TATA box were found in the upstream region of the soybean oleosin gene, which shows the functional oleosin promoter available. Homology comparison reveals that the soybean 24 kDa oleosin shares the highest identity with the soybean oleosin isoform A (U09118, GenBank), reaching to 98.4% in nucleotide. A soybean oleosinhirudin fusion gene driven by the oleosin promoter was constructed and inserted into plant binary expression vector. The intact tobacco plantlets were transformed by means of vacuum infiltration approach, with the Agrobacterium tumefaciens harboring the above vector. The transient correct expression of oleosin-hirudin fusion gene was identified by SDS/PAGE, western blotting and enterokinase treatment.     

Associations of Two Polymorphic Loci: A HinfⅠLocus of the Porcine Subunit C of Succinate Dehydrogenase Complex (SDHC) Gene and A MspⅠLocus of the Porcine Rod cGMP-Phosphodiesterase γ-Subunit (PDE6G) Gene

A Hinf I locus of the porcine subunit C of succinate dehydrogenase complex (SDHC) gene and a Msp I locus of the porcine rod cGMP-phosphodiesterase γ-subunit (PDE6G) gene had been reported before, but the association analysis between the different genotypes and the traits had not been done. 300 Large White × Meishan F2 pigs were used as experimental materials to performe the PCR-RFLP analysis and association analysis for the two loci, results revealed that the polymorphism of the porcine subunit C of succinate dehydrogenase complex (SDHC) gene was significantly associated with the traits which included the carcass length, the estimated lean meat percentage, the estimated backfat thickness at last rib, the estimated backfat thickness at last 3-4th rib, the fat meat weight, the fat meat percentage, the lean meat weight,the lean meat percentage, the ratio of lean meat to fat meat, the leaf fat weight, the backfat thickness at shoulder, the backfat thickness at thorax-Waist, the backfat thickness at 6-7th thorax and the average daily gain. Seven other traits, the meat color value (Biceps femoris, BF), the meat marbling (Biceps femoris, BF), the water moisture (Longissimus dorsi, LD),the bone weight, the bone percentage, the loin eye width and the loin eye area, were found to be significantly correlated with the polymorphism of the porcine rod cGMP-phosphodiesterase γ-subunit (PDE6G) gene. Based on these results, it is necessary to apply the two genes as candidate genes to marker assistant selection (MAS) in pig breeding.     

A Dominant Locus,qBSC-1, Controls β Subunit Content of Seed Storage Protein in Soybean (Glycine max (L.) Merri.)

Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F₂ segregated population and residual heterozygous lines (RHL) derived from the cross between Yangyandou (low level of β subunit) and Zhonghuang 13 (normal level of β subunit) were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1 (β subunit content), which was mapped to a region of 11.9 cM on chromosome 20 in F₂ population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome (Glyma 1.0, Williams 82), including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine (TT) insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection (MAS).     

Cloning and Expression Analysis of Sucrose Non-fermenting-1 Related Protein Kinase （SnRK） in Cucumis sativus L. Under Low Nitrogen Conditions

The sucrose non-fermenting-1 related protein kinase（SnRK）, whose expression is induced by kinds of hyperosmotic stresses, plays a key role in improving stress resistance of plants. In order to investigate the molecular mechanism of low nitrogen resistance in cucumber, the full-length cDNA of SnRK gene was cloned in this study. The result showed that SnRK gene was 1 548 bp in length, encoded 515 amino acids, and had more than 80% homology with other crops. The protein encoded by this gene was an unstable and hydrophilic protein with no transmembrane structure and no signal peptide. Under nitrogen-free conditions and low nitrogen conditions, the expression pattern analysis of SnRK gene showed that this gene was up-regulated and its expression increased and was significantly higher than the normal level as the nitrogen concentration decreased. In addition, the expression of SnRK gene was also inhibited in the high nitrogen level and was significantly lower than the normal level. The result of this study would help us understand the molecular mechanism of low nitrogen resistance in cucumber.     

Cloning and Functional Analysis of Lycopene ɛ-Cyclase (IbLCYe) Gene from Sweetpotato,Ipomoea batatas (L.)Lam.

This paper reported firstly successful cloning of lycopene ε-cyclase (IbLCYe) gene from sweetpotato, Ipomoea batatas (L.) Lam. Using rapid amplification of cDNA ends (RACE), IbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence of IbLCYe gene contained a 1 236 bp open reading frame (ORF) encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point (pI) of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD (flavinadenine dinucleotide)/NAD(P) (nicotinamide adenine dinucleotide phosphate)-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA of IbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco (cv. Wisconsin 38) expressing IbLCYe gene accumulated significantly more β-carotene compared to the untransformed control plants. These results showed that IbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.     

Variations on conserved signaling pathways in biocontrol and development:G protein and MAPK genes of Trichoderma. atroviride and T. virens

Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form …     

Expression Comparisons of Pathogenesis-Related (PR) Genes in Wheat in Response to Infection/Infestation by Fusarium,Yellow dwarf virus (YDV) Aphid-Transmitted and Hessian Fly

Expression profiles of ten pathogenesis-related （PR） genes during plant defense against Fusarium, Yellow dwarf virus （YDV） aphid-transmitted and Hessian fly （Hf） were compared temporally in both resistant and susceptible genotypes following pathogen infection or insect infestation. Quantitative real-time PCR （qRT-PCR） revealed that PR1, PR2, PR3, PR5, PR6, PR8, PR9, and PR15 appeared to be induced or suppressed independently in response to Fusarium, YDV aphid-transmitted or Hf during the interactions. The PR gene（s） essential to defense against one organism may play little or no role in defense against another pathogen or pest, suggesting the alternative mechanisms may be involved in different interactions of wheat- Fusarium, wheat-YDV aphid-transmitted and wheat-Hf. However, strong up- or down-regulation of PRl2 and PR14 encoding low molecular membrane acting protein, defensin and lipid transfer protein （LTP）, respectively, had been detected after either pathogen infection or insect infestation, therefore showed broad responses to pathogens and insects. It was postulated that low molecular proteins such as defensins and LTPs might play a role in the early stages of pathogenesis in the signaling process that informs plants about the attack from biotic stresses. In addition, a synergistic action between different PR genes might exist in plants to defense certain pathogens and insects on the basis of comprehensive expression profiling of various pathogenesis-related genes revealed by qRT-PCR in this study.     

Genetic Differentiation Analyses Based on mtDNA COⅡ Gene Sequences Among Different Geographic Populations of Aphis glycines(Hemiptera: Aphididae) in Northeast China

Aphis glycines(Hemiptera: Aphididae) is considered as a cosmopolitan pest of cultivated soybean, major diffi culties in its control measures may be due to its higher genetic diversity; however, the knowledge about population genetic diversity of this species is limited. This study aimed to represent the genetic differentiation among different geographic populations of soybean aphid in Northeast China. In order to investigate and assess the genetic diversity, genetic differentiation, molecular variance, population structure, ecological importance and evolutionary history of A. glycines, we sequenced a fragment of one protein-coding gene, the cytochrome c oxidaseⅡof mitochondrial DNA gene. The results showed that four haplotypes were defi ned among COⅡ gene of 180 sequences of soybean aphid in Northeast China including H1 shared by all the populations. Lower haplotype diversity(Hd=0.3590±0.0420) and nucleotide diversity(Pi=0.0012±0.0002) were observed and high gene flow was detected in every two populations, while most of the variation(80.81%) arose from variability within A. glycines from individuals. Low genetic differentiation and high gene fl ow(Nm=2.106) indicated a high migration rate between the populations, which might reveal that gene flow in different geographic populations did not affect by geographical distance. The phylogenetic tree and the haplotype network of A. glycines were obtained based on sequences of COⅡ gene, there were no signifi cant genealogical branches or clusters recognized in NJ tree, and no clear distribution, delineation of haplotypes were demonstrated in the haplotype network according to geographical location. This study rejected the vicariance hypothesis: geographic isolation could be a barrier and it restricted A. glycines gene fl ow among 10 populations.     

Effects of Temperature on Functional Response of Anagrus nilaparvatae Pang et Wang (Hymenoptera: Mymaridae) on the Eggs of Whitebacked Planthopper,Sogatella furcifera Horváth and Brown Planthopper,Nilaparvata lugens Stål

Understanding the temperature affecting parasitic efficiency is critical to succeed in utilizing parasitoid as natural enemy in pest management. Laboratory studies were carried out to determine the effects of temperature on parasitoid preference of female Anagrus nilaparvatae Pang et Wang (Hymenoptera: Mymaridae) to the eggs of whitebacked planthopper (WBPH), Sogatella furcifera Horváth and brown planthopper (BPH), Nilaparvata lugens Stål to build a composite model describing changes in parasitic response along a temperature gradient (18, 22, 26, 30, 34C). The results showed that attack responses of A. nilaparvatae on WBPH and BPH were the best described by a Type II functional response. The two parameters, attack rates (a) and handling times (T_h), of A. nilaparvatae to both eggs were influenced by the temperature. The maximum attack rates to WBPH (1.235) and BPH (1.049) were at 26 and 34C, respectively, and the shortest handling times to WBPH (0.063) and BPH (0.057) were at 30 and 26C, respectively. However, the optimal temperature for parasitic efficiency of A. nilaparvatae to WBPH and BPH eggs was both at 26C, which showed that the present microclimate temperature of the habitat in the paddyfield was beneficial to A. nilaparvatae and indicated that parasitic efficiency of A. nilaparvatae would be impaired by global warming.