Nitrogen (N) enrichment is expected to induce a greater phosphorus (P) limitation, despite the acceleration of soil P cycling.  However, the changing patterns in plant P and soil available P after N enrichment, and their regulatory mechanisms, remain poorly understood in alpine meadows.  Here, we conducted a field experiment with four N addition rates (0, 5, 10, and 15 g N m^–2 yr^–1) in an alpine meadow, and investigated the P in plants, microorganisms, and soil to determine their patterns of change after short-term N addition.  Our results showed that N addition significantly increased plant biomass, and the plant P pool showed a non-linear response to the N addition gradient.  Soil available P initially increased and then declined with increasing N addition, whereas the occluded inorganic P decreased markedly.  The critical factors for soil available P varied with different N addition rates.  At lower N addition levels (0 and 5 g N m^–2 yr^–1), soil acidification facilitated the mobilization of occluded inorganic P to increase soil available P.  Conversely, at higher N addition levels (10 and 15 g N m^–2 yr^–1), the elevated soil microbial biomass P intensified the competition with plants for soil P, leading to a decline in soil available P.  This study highlights the non-linear responses of the plant P pool and soil available P concentration to N addition rates.  These responses suggest the need for developing ecosystem models to assess different effects of increasing N rates, which would enable more accurate predictions of the plant P supply and soil P cycling under N enrichment.

Cre/loxP, a site-specific recombination system, has been widely used for various purposes, including chromosomal translocations, generation of marker-free transgenic plants, tissue-specific activation of a reporter gene and efficient heterologous gene expression in plants.  However, stable or transient expression of Cre recombinase in plants can cause chlorosis or necrosis.  Here, we describe a modified Cre/loxP recombination system using a DNA fragment flanked with loxP sites in the same orientation in which necrosis induced by Cre recombinase in Nicotiana benthamiana leaves was alleviated.  The modified system was successfully used to create functional GFP-tagged pepper mild mottle virus (PMMoV) and a chimeric virus with coat protein (CP) substitution assembled from separate pro-vector modules.  Our results provide a new strategy and flexible technique to construct chimeric virus and infectious clones for plant viruses with large genomes.

Founder parents have contributed significantly to the improvement of wheat breeding and production.  In order to investigate the genetic characteristics of founder parents and widely planted cultivars, Mazhamai (M), Biyumai (B) and six sibling lines (BM1–6) derived from the cross M×B were phenotyped for eight yield-related traits over multiple years and locations and genotyped using the the wheat 90K single nucleotide polymorphism (SNP) assay.  BM4 has been used as a founder parent, and BM1 has been widely planted, whereas BM2, 3, 5, and 6 have not been used extensively for breeding or planting in China.  Phenotypic comparisons revealed that BM4 and BM1 displayed a better overall performance than the other sibling lines.  BM1 showed higher thousand-grain weight than BM4, whereas BM4 exhibited lower coefficient of variation for most of the yield-related traits across different years and locations, indicating that BM4 was widely adaptable and more stable in different environments.  SNP analysis revealed that BM4 and BM1 inherited similar proportions of the M genome but are dissimilar to BM2, 3, 5, and 6.  Both BM1 and BM4 have specific alleles that differ from the other BM lines, and most of these alleles are concentrated in specific chromosomal regions that are found to associate with favorable QTLs, these SNPs and their surrounding regions may carry the genetic determinants important for the superior performance of the two lines.  But BM4 has more genetic diversity than BM1 with more specific alleles and pleiotropic regions, indicating that the genome of BM4 may be more complex than the other sibling lines and has more favorable gene resources.  Our results provide valuable information that can be used to select elite parents for wheat and self-pollinating crop breeding.

   Grain number per spike (GNPS) is a major factor in wheat yield breeding.  A new wheat germplasm Pubing 3504 shows superior features in spike traits.  To elucidate the genetic basis of spike and yield related traits in Pubing 3504, 282 F2:3 families were generated from the cross Pubing 3504×Jing 4839, and seven spike and yield related traits, including GNPS, spike length (SL), kernel number per spikelet (KPS), spikelet number per spike (SNS), thousand-grain weight (TGW), spike number per plant (SNP), and plant height (HT) were investigated.  Correlation analysis indicated significant positive correlations between GNPS and spike-related traits, including KPS, SNS, and SL, especially KPS.  A genetic map was constructed using 190 polymorphic simple sequence repeat (SSR), expressed sequence tag (EST)-SSR, and sequence-tagged-site (STS) markers.  For the seven traits measured, a total of 37 quantitative trait loci (QTLs) in a single-environment analysis and 25 QTLs in a joint-environment analysis were detected.  Additive effects of 70.3% (in a single environment) and 57.6% (in a joint environment) of the QTLs were positively contributed by Pubing 3504 alleles.  Five important genomic regions on chromosomes 1A, 4A, 4B, 2D, and 4D could be stably detected in different environments.  Among these regions, the marker interval Xmag834–Xbarc83 on the short arm of chromosome 1A was a novel important genomic region that included QTLs controlling GNPS, KPS, SNS, TGW, and SNP with stable environmental repeatability.  This genomic region can improve the spike trait and may play a key role in improving wheat yield in the future.  We deduced that this genomic region was vital to the high GNPS of Pubing 3504.

    The enzyme Δ³,Δ²-dienoyl-CoA isomerase (ECI1) plays a crucial role in the mitochondrial β-oxidation of fatty acids with a double-bond in odd and even positions. The ECI1 gene might be a qualified candidate for studies pertaining to lipid deposition and meat quality in swine. In the present study, ECI1 cDNA of the Tibetan pig was obtained by in silico cloning and verified by PCR analysis. Single-nucleotide polymorphisms (SNPs) of ECI1 were screened by PCR-sequencing and genotypes of those SNPs were tested by PCR-restriction fragment length polymorphism (PCR-RFLP) in Diannan small-ear pigs (DSP, n=40), Tibetan pigs (TP, n=60) and Yorkshire pigs (YP, n=30). The expression levels of ECI1 were analyzed by real-time quantitative PCR and Western blotting in tissues of the liver, backfat, and longissimus dorsi (LD) muscle of DSP (n=8), TP (n=8) and YP (n=8). Single factor linear correlation analysis was applied separately for each breed to evaluate correlations between ECI1 gene expression in the LD muscle and intramuscular fat (IMF) content. We obtained an ECI1 gene length of 1 401 bp from the cDNA that contained a full coding region of 909 bp. Three novel SNPs (g.42425337G>A; g.42424666A>G; and g.42422755A>G) were detected, and only g.42424666A>G exhibited three genotypes among the three breeds. The ECI1 expression levels in the LD muscle of DSP and TP were significantly higher than that of YP (P<0.05). Moreover, TP had the highest ECI1 expression in backfat (P<0.01), and a positive correlation was observed between gene expression and IMF content. The results suggest that differences in ECI1 gene expression might be related to lipid deposition and meat quality in pig.

The wheat grain number per spike (GNPS) is a major yield-limiting factor in wheat-breeding programs. Germplasms with a high GNPS are therefore valuable for increasing wheat yield potential. To investigate the molecular characteristics of young spike development in large-spike wheat germplasms with high GNPS, we performed gene and protein expression profiling analysis with three high-GNPS wheat lines (Pubing 3228, Pubing 3504 and 4844-12) and one low-GNPS control variety (Fukuho). The phenotypic data for the spikes in two growth seasons showed that the GNPS of the three large-spike wheat lines were significantly higher than that of the Fukuho control line. The Affymetrix wheat chip and isobaric tags for relative and absolute quantitation-tandam mass spectrometry (iTRAQ-MS/MS) technology were employed for gene and protein expression profiling analyses of young spike development, respectively, at the floret primordia differentiation stage. A total of 598 differentially expressed transcripts (270 up-regulated and 328 down-regulated) and 280 proteins (122 up- regulated and 158 down-regulated) were identified in the three high-GNPS lines compared with the control line. We found that the expression of some floral development-related genes, including Wknox1b, the AP2 domain protein kinase and the transcription factor HUA2, were up-regulated in the high-GNPS lines. The expression of the SHEPHERD (SHD) gene was up-regulated at both the transcript and protein levels. Overall, these results suggest that multiple regulatory pathways, including the CLAVATA pathway and the meristem-maintaining KNOX protein pathway, take part in the development of the high-GNPS phenotype in our wheat germplasms.

Maintenance and management of genetic diversity of farm animal genetic resources (AnGR) is very important for biological, socioeconomical and cultural significance. The core concern of conservation for farm AnGR is the retention of genetic diversity of conserved populations in a long-term perspective. However, numerous factors may affect evolution of genetic diversity of a conserved population. Among those factors, the genetic architecture of conserved populations is little considered in current conservation strategies. In this study, we investigated the dynamic changes of genetic diversity of conserved populations with two scenarios on initial genetic architectures by computer simulation in which thirty polymorphic microsatellite loci were chosen to represent genetic architecture of the populations with observed heterozygosity (Ho) and expected heterozygosity (He), observed and mean effective number of alleles (Ao and Ae), number of polymorphic loci (NP) and the percentage of polymorphic loci (PP), number of rare alleles (RA) and number of non-rich polymorphic loci (NRP) as the estimates of genetic diversity. The two scenarios on genetic architecture were taken into account, namely, one conserved population with same allele frequency (AS) and another one with actual allele frequency (AA). The results showed that the magnitude of loss of genetic diversity is associated with genetic architecture of initial conserved population, the amplitude of genetic diversity decline in the context AS was more narrow extent than those in context AA, the ranges of decline of Ho and Ao were about 4 and 2 times in AA compared with that in AS, respectively, the occurrence of first monomorphic locus and the time of change of measure NP in scenario AA is 20 generations and 23 generations earlier than that in scenario AS, respectively. Additionally, we found that NRP, a novel measure proposed by our research group, was a proper estimate for monitoring the evolution of genetic diversity in a closed conserved population. Our study suggested that current managements of conserved populations should emphasize on initial genetic architecture in order to make an effective and feasible conservation scheme.