Previous Aphid Infestation Induces Different Expression Profiles of Genes Associated with Hormone-Dependent Responses in Near-Isogenic Winter Wheat Lines.

Hormone-dependent responses in host plants induced by herbivore infestation have species-specific effects. This study focused on determining the relative expression profiles of the genes associated with hormone-dependent pathways in two near-isogenic wheat lines when attacked by cereal aphids. Infestation with Rhopalosiphum padi Linnaeus (Hemiptera: Aphididae) and/or Sitobion avenae Fabricius (Hemiptera: Aphididae) significantly upregulated the expression of marker genes related to the salicylic acid (SA)- and jasmonic acid (JA)-dependent pathways in the tested lines. In the resistant line 35-E4, previous infestation with R. padi significantly increased the relative expression of plant pathogenesis-related protein 1 at all sampling times but did not have a significant effect on the expression of the phenylalanine ammonia-lyase (PAL) gene. In addition, the expression levels of the lipoxygenase (LOX) and allene oxide synthase (AOS) genes immediately increased after the aphid attack. In susceptible line 35-A20, infestation with either R. padi or S. avenae led to significantly increased expression levels of the AOS and PAL genes. Moreover, sequential aphid infestation induced higher expression of AOS compared with a single-species aphid infestation, whereas the expression of the PAL gene was antagonistically affected by sequential aphid infestation. Overall, these results showed that aphid infestation induced SA- and JA-dependent responses in host plants. However, the expression profiles of these genes in resistant and susceptible host lines were significantly different.

Direct Exposure of Wheat Seeds to High-Voltage Electrostatic Fields Adversely Affects the Performance of Sitobion avenae (Hemiptera: Aphididae).

Exposing seeds to a high-voltage electrostatic field (HVEF) may influence the performance of herbivores and improve the germination rate by inducing biological and physiological changes in plants. In the current study, an age-stage, two-sex life table was established to evaluate the effects from directly exposing seeds to HVEFs on the performance of apterous Sitobion avenae (Fabricius) reared on winter wheat. We treated the wheat seeds by exposing them to an HVEF for 20 min at three intensities: 2, 4, or 6 KV/cm. Controls received no treatment (0 KV/cm). The results indicated that the parameters of the net reproductive rate (R0), the intrinsic rate of increase (r), and the mean generation span (T) of S. avenae were significantly reduced by HVEFs through multiple generations. In addition, the age-specific survival rate (lx) and comparison with the results of a Weibull equation analysis suggested that S. avenae had the shortest life span when seeds were exposed to an HVEF at 4 KV/cm. Overall, these findings indicated that direct exposure of wheat seeds to an HVEF at 4 KV/cm could adversely affect the performance of S. avenae.

Resistance of Wheat Accessions to the English Grain Aphid Sitobion avenae.

The English grain aphid, Sitobion avenae, is a major pest species of wheat crops; however, certain varieties may have stronger resistance to infestation than others. Here, we investigated 3 classical resistance mechanisms (antixenosis, antibiosis, and tolerance) by 14 wheat varieties/lines to S. avenae under laboratory and field conditions. Under laboratory conditions, alatae given the choice between 2 wheat varieties, strongly discriminated against certain varieties. Specifically, the 'Amigo' variety had the lowest palatability to S. avenae alatae of all varieties. 'Tm' (Triticum monococcum), 'Astron,' 'Xanthus,' 'Ww2730,' and 'Batis' varieties also had lower palatability than other varieties. Thus, these accessions may use antibiosis as the resistant mechanism. In contrast, under field conditions, there were no significant differences in the number of alatae detected on the 14 wheat varieties. One synthetic line (98-10-30, a cross between of Triticum aestivum (var. Chris) and Triticum turgidum (var. durum) hybridization) had low aphid numbers but high yield loss, indicating that it has high antibiosis, but poor tolerance. In comparison, 'Amigo,' 'Xiaoyan22,' and some '186Tm' samples had high aphid numbers but low yield loss rates, indicating they have low antibiosis, but good tolerance. Aphid population size and wheat yield loss rates greatly varied in different fields and years for '98-10-35,' 'Xiaoyan22,' 'Tp,' 'Tam200,' 'PI high,' and other '186Tm' samples, which were hybrid offspring of T. aestivum and wheat related species. Thus, these germplasm should be considered for use in future studies. Overall, S. avenae is best adapted to 'Xinong1376,' because it was the most palatable variety, with the greatest yield loss rates of all 14 wheat varieties. However, individual varieties/lines influenced aphid populations differently in different years. Therefore, we strongly recommend a combination of laboratory and long-term field experiments in targeted planting regions to identify varieties/lines that consistently show high resistance to S. avenae infestation.

Testing the fecundity advantage hypothesis with Sitobion avenae, Rhopalosiphum padi, and Schizaphis graminum (Hemiptera: Aphididae) feeding on ten wheat accessions.

The fecundity advantage hypothesis suggests that females with a large body size produce more offspring than smaller females. We tested this hypothesis by exploring the correlations between life-history traits of three aphid species feeding on ten wheat accessions at three levels of analysis with respect to the host plant: overall, inter-accession, and intra-accession. We found that fecundity was significantly correlated with mean relative growth rate (MRGR), weight gain, and development time, and that the faster aphid develops the greater body and fecundity, depending on aphid species, wheat accession, and analyses level. Larger aphids of all three species produced more offspring overall; this held true for Sitobion avenae and Schizaphis graminum at the inter-accession level, and for S. avenae, Rhopalosiphum padi, and S. graminum for three, five, and eight accessions respectively at the intra-accession level. Only one correlation, between intrinsic rates of natural increase (rm) and MRGR, was significant for all aphid species at all three analysis levels. A more accurate statement of the fecundity advantage hypothesis is that cereal aphids with greater MRGR generally maintain higher rm on wheat. Our results also provide a method for exploring relationships between individual life-history traits and population dynamics for insects on host plants.

Meiotically asynapsis-induced aneuploidy in autopolyploid Arabidopsis thaliana.

The patterns of homologue segregation are the basis for euploidy or aneuploidy formation in diploids and allo-/auto-polyploids. Homologue segregation in diploids resembles that in allopolyploids during meiosis; however, meiotic chromosome behavior in autopolyploids is complicated by multiplication of homologous chromosome components. Obviously, loss of single chromosomes (or segmented chromosomes) frequently leads to abortion of reproductive gametes in diploids and allopolyploids. In contrast, the consequence of chromosome loss in autopolyploids is effortlessly compensated for by the presence of multiplied chromosome complements. Here, we use the meiotically asynaptic gene asy1, in combination with polyploidization, to elucidate aneuploidy formation in autotetraploid Arabidopsis. The results indicate that, due to homologous asynapsis in meiotic prophase I, retarded chromosome losses could induce aneuploidy during gametogenesis in autotetraploid asy1. The severe loss of individual chromosomes probably reaches the haploid genome among selfed or backcrossed progeny, leading to stochastic chromosome loss in Arabidopsis. Reciprocal crosses of autotetraploid asy1 with wild-type prove a pathway of duoparental transmission of aneuploidy (hypoploidy and hyperploidy). Viable hypoploids over-transmit via male gametes; conversely, viable hyperploids transmit mainly in female gametogenesis. This result suggests a more stringent maternal restriction of ploidy transmission in autopolyploid Arabidopsis.

[Two-dimensional electrophoresis on anther total proteins of male sterility induced by chemical hybridizing agent and heredity type in wheat].

The isonucleus-allocytoplasmic materials ms(S)-Xinong 1376, (A)-Xinong 1376, and ms(A)-Xinong 1376 were used to investigate the protein expression profile in mononucleated anaphase anthers of wheat (Triticum aestivum L.) after silver staining and analysis by PDQuest software. About 320 to 350 distinct protein spots were detected reproducibly. The results indicated that there were some difference in the protein quantity between the two sterile lines and the maintainer line. Several specific spots were present in the maps of two sterile lines, but were absent in the maintainer line. The two sterile lines had their own specific expression protein spots with different sterile mechanism. We compared the three maps and concluded that the sterile lines may inhibit related protein expression, and turned on some proteins related to male sterility, which disturbs the normal metabolism of substances and energy in anthers and leads to the occurrence of male-sterility.

[An efficient method for isolation of mitochondrial DNA in wheat].

An efficient method for isolation of mitochondrial DNA (mtDNA) from etiolated tissues of wheat was developed. The protocol consists of mitochondria isolation with differential centrifugation, Dnase I treatment, lysis with SDS and proteinase K, removing protein by TE-saturated phenol/chloroform extraction and a final RNase A treatment for obtaining mtDNA. The mtDNA samples were tested using spectrophotometry and agarose gel electrophoresis. It was proved that the mtDNA isolated by this method not only have the high yield but also structural complete, and contains no impurities, such as nuclear DNA, RNA and protein. The result showed that this high quality mtDNA can be successfully used in PCR and other genetic studies. In addition, it was found that adjusting the lysis temperature has a noticeable effect on the mtDNA yield.

Induced expression of DREB transcriptional factor and study on its physiological effects of drought tolerance in transgenic wheat.

Expression vector pBAC128F, which carries DREB transcriptional factor gene driven by drought inducing promoter rd29B and bar gene driven by CaMV 35S promoter and maize Adh1 gene first intron, was transferred into the explants of immature inflorescence and immature embryos of hexaploid winter wheat cv. 8901, 5-98, 99-92 and 104 by particle bombardment. More than 70 resistant transgenic plants were obtained. Genomic PCR and RNA dot blotting analyses showed that DREB gene had been integrated into wheat genome of the transgenic plants (T0 and T1) and was well expressed in offspring seed of different transgenic lines. The content of proline in leaves and seeds of T2 transgenic lines was analyzed. Among 16 tested transgenic lines, 10 transgenic lines exhibited more than two fold of proline level in leaves as compared with CK plants. Under drought condition, after stopping water for 15 days the leaves of transgenic lines were still green, while CK were faded. After rewatering for 10 days, the leaves of transgenic lines maintained their green, while all CK plants were dead. Our research suggested that introducing a novel DREB transcriptional factor into wheat is an effective way to improve its drought-tolerance ability.

[Identification of wheat and barley hybrid offspring by GISH and PAGE].

Genomic in situ hybridization (GISH) and polyacrylamide gel electrophoresis (PAGE) of wheat seed were used to identify the barley chromosome in offspring of wheat and barley hybrid. A sires of alien addition lines, alien substitution lines and translocation lines were detected by GISH. WBA984 and WBA9812 were alien addition lines, WBS0215 and WBS0264 were alien substitution lines,and WBT02125 and WBT02183 were translocation lines in chromosome terminal. SDS-PAGE analysis and A-PAGE patterns indicated that WBA9812 were 5H alien addition line, WBS0264 was 1 B/5H alien substitution line, and WBT02125 was 1 BL/5HL translocation line.

[Studies on application of APAGE in male sterile wheat with alloplamic cytoplasm].

Four male sterile wheat with Aeglops kotschyi, Ae. variabilis, Ae. ventricosa and Ae. bicornis cytoplasm, were crossed with some wheat materials, and from the crosses a series of maintenance lines were selected. The APAGE(acid polyacrylamide gel electrophoresis) analysis indicated that most maintenance lines possessed the gliadin marker Gld1B3 of 1BL/1RS translocation chromosome. The root tip chromosomes of this maintenance lines had two satellite chromosomes, but other maintenance lines without Gld1B3 marker had the same four satellite chromosomes as other common wheat. Three male sterile wheats with Aeglops kotschyi, Ae. variabilis, Ae. ventricosa cytoplasm had the same fertility, but the male sterile wheat with Ae. bicornis cytoplasm could produce male sterile. Meantime, the paper discussed application of APAGE technology in breeding of non-1BL/1RS male sterile wheat.