Insect pest and weeds are two major problems for forage and turf grasses. In this study, scarab larvae- and herbicideresistant transgenic perennial ryegrass (Lolium perenne L.) was obtained by transforming it with cry and bar genes simultaneously via the Agrobacterium-mediated method. To optimize the callus induction and plant regeneration conditions, various concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine were assayed. The transformation efficiencies of different Agrobacterium suspension media, used during Agrobacterium-mediated transformation, were compared. Then, plasmids of pCAMBIA3301 containing cry gene (cry8Ca2 or cry8Ga) and bar gene, driven by ubiquitin promoter, were transformed into perennial ryegrass. The transformants were generated and confirmed by both Southern hybridization analysis and Western hybridization analysis. Further, the resistance of transgenic perennial ryegrass plants to scarab larvae and herbicide were analyzed. After 30 d of co-cultivation with scarab larvae, the damage to the root system of transgenic plants was less than that of non-transgenic control plants. Additionally, the leaves of transgenic plants were resistant to Basta®, while leaves of the wild plants wilted after Basta® spraying. These results show that cry gene and bar gene were successfully transferred into perennial ryegrass by the Agrobactgerium-mediated method, and convey resistance to scarab larvae and herbicide in transgenic perennial ryegrass plants.

To evaluate the month-wise prevalence of subclinical mastitis (SM) and its relations with climatic temperature and humidity, and to provide references for control and prevention of mastitis in dairy cows in Guangdong, China, California mastitis test method was used to determine the monthly prevalence of SM (MPSM, %) of lactating Holstein cows from 11 dairy farms in Guangdong Province, China, including about 516 cows per monthly examination. The average MPSM on a cow and a quarter basis were 30 and 13% respectively, and there was a positive linear correlation between monthly mean air temperature (MT, °C) and MPSM on a cow basis (r=0.763, P=0.004) or a quarter basis (r=0.577, P=0.049), but there were no correlations between MT and MPSM on a cow or a quarter basis (P>0.05). There was a shift in MPSM trend that the highest MPSM (38.4%) was not for the hottest July with MT at 30.8°C, but for the later September at 27.9°C. The farms need to develop new housing conditions to control cowshed temperatures in the hot season to reduce the mastitis prevalences in the posthot months.

Microbiota in the gastrointestinal tract (GIT) of piglets during weaning transition can experience a sharp change which could result in growth reduction and diarrhea of weaned piglets. Dietary manipulations can play an important role in attenuating such changes caused by weaning stress. Therefore, ileal and colonic contents of weaned piglets were used as inocula, mannan oligosaccharide (MOS) or sugar beet pulp (SBP) was supplied as single energy sources to investigate effects of MOS or SBP on the shifts of gastro-intestinal microflora and lactobacilli populations. The universal bacteriaand lactobacilli-specific PCR/denaturing gradient gel electrophoresis (DGGE), cloning and sequencing techniques were used. DGGE profiles of the universal bacteria showed that great changes were found in the position, numbers and intensity of dominant bands after fermentation. The similarity of bacterial community between ileum and colon was increased to 85-97% by MOS or SBP treatment after fermentation from the similarity with 20% before fermentation. MOS treatment significantly increased the bacterial diversity and band number in both ileal and colonic fermentation (P<0.05). SBP treatment significantly increased the bacterial diversity and band number in colon (P<0.05). It implies that some species were enriched by the addition of MOS or SBP to increase the similarity and diversity of bacterial community in weaned piglets. Five specific bands appearing in MOS or SBP treatment group after fermentation were cloned and sequenced, the changes of species related to Prevotella and Ruminococcus were observed. Two bands related to uncultured bacterium with 98% similarity were detected by MOS or SBP treatment. However, there were no effects on the similarity, diversity index and lactobacilli species revealed by MOS or SBP treatment. These results imply that MOS or SBP could have beneficial effects on the weaning piglets by stablizing microbiota in the GIT microflora.

Five commonly-used reference genes: ACT (actin), UBE (ubiquitin-conjugating enzyme), RPL2 (ribosomal protein L2), BRP II (RNA polymerase II subunit), and NADH (nicotinamide adenine dinucleotide) were examined using geNorm software as reference genes for RT-qPCR. Among the tested reference genes, ACT and UBE were the most stable in all samples. In parallel, expression analysis of nitrilases in Brassica juncea var. tumida, was performed to preliminarily investigate the molecular interactions between nitrilase and clubroot development at 10, 15, 20, 25, 30, and 40 d postinoculation (dpi) with a suspension of resting spores of Plasmodiophora brassicae. The results showed that different gene expressions of nitrilases were regulated during the initial periods of clubroot development. The expression level of BjNIT1 increased sharply from 20 to 40 dpi in infected roots while there were no remarkable changes in healthy roots. From 15 to 30 dpi, the expression levels of BjNIT2 and BjNIT4 in infected roots were lower than those in non-infected roots. Finally, BjNIT2 in treatment was down approximately to control at 40 dpi. Our results suggest that BjNIT1, which promoted overproductions of auxin, might be involved in P. brassicae infection of B. juncea.

Abstract To assess the effects of creatine pyruvate (Cr-Pyr) on lipid and protein metabolism in broiler chickens, a total of 400 1-day-old male birds (Aconred) were randomly allocated to four groups, with each group replicating four times and each replicate involving 25 birds. The broilers were provided with a commercial diet supplemented with Cr-Pyr at 0, 1, 5, or 10% of the diet, respectively, for a period of 3 wk ad libitum (from 22 to 42 d). In the present study, body weight (BW) and average daily gain (ADG) of broilers decreased in 10% Cr-Pyr group ( P<0.01), whereas the relative leg and pectoral muscle weights were significantly higher than they were in the control group (P<0.05). 5 or 10% Cr-Pyr of diets decreased the abdominal fat rate (AFR, abdominal fat/live weight) of the broilers. The serum or hepatic triglyceride (TG) concentrations were significantly lower in the 5 and 10% groups (P<0.01). In contrast, Cr-Pyr caused a marked increase in the serum non-esterified fatty acid (NEFA), high-density lipoprotein cholesterol (HDL-C) and creatine kinase (CK) concentrations (P<0.01). Supplementation with Cr-Pyr (5 and 10%) in the diet also increased glucagons (GLU), insulin (INS) or leptin (LEP) contents (P<0.01). The expression of hepatic peroxisomal proliferators-activated receptor a (PPAR-a) and carnitine palmitoyl transferase-I (CPT-I), muscle insulin-like growth factor I (IGF-I) were significantly elevated and myostatin mRNA level was reduced in the 5 and 10% groups (P<0.05). It was found that supplementation with 5% Cr-Pyr improves both lipid and protein metabolism by regulating various metabolic parameters of broilers, while not adversely affects growth performance in broiler chickens.

Abstract This research was aimed to develop AFLP markers co-segregated with gene Lr24 and validate the using for marker assisted selection (MAS). An F 2 population developed from the cross between the resistant line TcLr24 and the susceptible line Thatcher was tested for resistance to the Puccinia triticina races BGQQ and SHRT using for genetic analysis and molecular marker. A total of 224 AFLP primer combinations were used to test the resistant and susceptible parents, as well as the resistant bulk and the susceptible bulk. Four AFLP markers, P-AGA/M-CTT 289 bp, P-AGC/M-CAC 188 bp, P-AGC/M-CAC 162 bp and P-ACG/M-CGC 239 bp were co-segregated with Lr24. The AFLP fragment from the primer combination P-ACG/M-CGC was cloned, sequenced and converted into a STS marker named as ASTS212. Thatcher backgrounded NILs and 115 varieties were examined by using this STS marker and the marker SCS1302 607 developed by Gupta. 5R615, 5R616, 1R13, and 1R17 were identified and validated to contain gene Lr24. The marker is dominant and may be useful in identification the resistance gene Lr24 in wheat and wheat breeding programs.

Abstract Thermal degradation kinetics of anthocyanins and visual color (Hunter a* value) of blood orange juice were studied at selected temperatures (70-90°C). Results indicated that both the thermal degradation of anthocyanin and visual color all followed first-order reaction kinetics, and they could be expressed by Arrhenius equation. The activation energy values for the anthocyanins degradation and visual color degradation were 55.81 and 47.51 kJ mol −1, respectively. The linear relationship between visual color and anthocyanin content was obtained. Furthermore, during thermal processing of blood orange juice, the formulas about the linear relationships showed no significant difference at selected temperatures. So, the relationships between visual color and anthocyanins content during thermal processing at selected temperatures could be described by the same equation: a*/a 0*=0.559(C/C 0)+0.43. It might be inferred that visual color measured instantaneously by tristimulus colorimeters for on-line quality control, could be used to predict the anthocyanins degradation during thermal processing of blood orange juice.

Abstract We investigated the size distribution of water-stable aggregates and the soil carbon, nitrogen and phosphorus concentration over aggregate size fractions based on a long-term (1990-2006) fertilization experiment in a reddish paddy soil. The results showed that the largest water-stable aggregate (WSA) (>5 mm) and the smallest WSA (<0.25 mm) took up the first largest proportion (38.3%) and the second largest proportion (23.3%), respectively. Application of organic materials increased the proportion of the large WSA (>2 mm) and decreased the proportion of the small WSA (<1 mm), resulting in an increase in the mean weight diameter of WSA, whereas application of chemical fertilizer had little effect. Application of organic materials, especially combined with chemical fertilizers, increased total carbon, nitrogen and phosphorus concentrations in all sizes of WSA, and total carbon, nitrogen and phosphorus were prone to concentrate in the large WSA. Further more, application of organic materials improved the supply effectiveness of available phosphorus, whereas had little influence on the labile carbon in WSA. Application of chemical fertilizers improved concentrations of total and available phosphorus in all sizes of WSA, whereas had little influence on total carbon and nitrogen contents. Economical fertilization model maintained the soil fertility when compared with full dose of chemical fertilizers, indicating that using organic materials could reduce chemical fertilizers by about one third.