Canola (Brassica napus) germplasm shows variable allelopathic effects against annual ryegrass (Lolium rigidum)

Aims

The allelopathic activity of canola (Brassica napus) germplasm was investigated using the important Australian weed, annual ryegrass (Lolium rigidum) as the target species.

Methods

Three different canola plant densities (10, 20, and 30 seedlings/beaker) of each of 70 world-wide genotypes were tested in vitro in close proximity to annual ryegrass seedlings.

Results

The allelopathic activity of canola, as measured by reduction in annual ryegrass root and shoot growth, increased with canola crop seedling densities. Density did not consistently influence shoot length of annual ryegrass. Greater shoot length suppression was observed in genotype cv. Rivette and BLN3343CO0402. The Australian genotype cv. Av-opal and the breeding line Pak85388-502 suppressed root length of ryegrass more than other genotypes, even at low densities. At the lowest density, the least allelopathic genotypes were cv. Barossa and cv. Cescaljarni-repka, although they became more allelopathic at higher density. An overall inhibition index was calculated to rank each of the canola genotypes. There were significant differences between canola genotypes in their ability to inhibit root and shoot growth in ryegrass.

Conclusion

Considerable genetic variation exists among canola genotypes for their allelopathic effects on annual ryegrass. Further investigation is required to determine the allelopathic mechanisms, particularly to identify the responsible allelochemical(s) and the gene(s) controlling the trait. This research suggests that highly allelopathic canola genotypes can be potential for controlling weeds such as annual ryegrass in integrated weed management programs.     

Molecular cloning, identification, and chromosomal localization of two MADS box genes in peach （Prunus persica）

MADS box proteins play an important role in floral development. To find genes involved in the floral transition of Prunus species, cDNAs for two MADS box genes, PpMADS1 and PpMADS10, were cloned using degenerate primers and 5'- and 3'- RACE based on the sequence database of P. persica and P. dulcis. The full length of PpMADS1 eDNA is 1, 071bp containing an open reading frame (ORF) of 717bp and coding for a polypeptide of 238 amino acid residues. The full length of PpMADS10 cDNA is 937bp containing an ORF of 633bp and coding for a polypeptide of 210 amino acid residues. Sequence comparison revealed that PpMADS1 and PpMADS10 were highly homologous to genes AP1 and PI in Arabidopsis, respectively. Phylogenetic analysis indicated that PpMADS1 belongs to the euAP1 clade of class A, and PpMADS10 is a member of GLO/PI clade of class B. RT-PCR analysis showed that PpMADS1 was expressed in sepal, petal, carpel, and fruit, which was slightly different from the expression pattern of AP1; PpMADS10 was expressed in petal and stamen, which shared the same expression pattern as PI. Using selective mapping strategy, PpMADS1 was assigned onto the Bin 1:50 on the G1 linkage group between the markers MCO44 and TSA2, and PpMADS10 onto the Bin 1:73 on the same linkage group between the markers Lap-1 and FGA8. Our results provided the basis for further dissection of the two MADS box gene function.     

Multiple infection of TT virus (TTV) with different genotypes in Japanese hemophiliacs

To clarify the persistent TT virus (TTV) infection, we studied a possibility of multiple TTV infection by genotype analysis of isolated TTV obtained from seven Japanese hemophiliacs. The nucleotide sequences including 222 bp in the open reading frame 1 (ORF1) region of 10 TTV isolates from each patient were analyzed and classified into various TTV genotypes such as G1 to G6 by phylogenetic analysis using a N-J method. Multiple TTV genotypes were observed in all the hemophiliacs: three different TTV genotypes were found in three patients, whereas four different TTV genotypes were observed in the other three patients. The remaining patient was also infected with TTV of five different genotypes. Moreover, new TTV genotypes were found in these seven patients and tentatively designated as G7. The present findings indicate that multiple TTV infection with different TTV genotypes has occurred in Japanese hemophiliacs. They also provide valuable information to understand persistent TTV infection.     

Effect of P sources on growth, P accumulation and activities of phytase and acid phosphatases in two cultivars of annual ryegrass (Lolium multiflorum L.).

Recurrent application of animal manure to the soil often results in accumulation of phosphorus (P) in the soil over time. Use of temperate forages like Lolium multiflorum capable of extracting excess P from manure impacted soil is an attractive strategy for P phytoremediation. Two genotypes of L. multiflorum, 'Gulf and Marshall' were grown in soil and hydroponic media containing various concentrations of poultry manure and their P accumulation potential was determined. A decline in the biomass with an increase in manure concentration beyond 10 g kg(-1) soil in Gulf and 25 g kg(-1) soil in Marshall was noticed. Gulf grass accumulated more P content (7 g kg(-1) dry weight) as compared to Marshall (6 g kg(-1) dry weight) in both roots and shoots. Maximum shoot P content was observed in the soil amended with 10 g poultry manure, while root P was highest at the concentration of 50 g poultry manure kg(-1) in the soil. Both cultivars yielded the highest biomass when grown in the presence of 10 g poultry manure in modified Hoagland's media. Presence of chelators in the media did not produce any noticeable effect on P accumulation in either grass and the biomass was appreciably enhanced by all concentrations of the chelators. Gulf and Marshall ryegrass seedlings were grown hydroponically in various poultry manure fractions. Both phytase and acid phosphatase (APase) enzyme activities in the root increased substantially in response to P-sufficient condition. In the presence of various poultry manure fractions, an intermediate level of both enzymes was measured compared to the P-sufficient condition, while the lowest enzyme activity was observed in the absence of any P source in the media. The level of APase and phytase activities was more or less the same in the two grasses under various growth conditions. An additional APase isoform was induced specifically in response to P-starvation from the two grass cultivars. Phytase and APase assays carried out in the P-starved and P-replenished grass seedlings further confirmed that during P deficiency, the enzyme activity was lowest and results of PAGE indicated that an APase isoform was induced under P-starvation.     

Molecular cloning and tissue-specific expression of two cDNAs encoding polyketide synthases from Hypericum perforatum

Two previously uncharacterized cDNAs encoding for polyketide synthases (PKSs), designated as HpPKS1 and HpPKS2, were isolated from Hypericum perforatum. The full-length HpPKS1 was 1573bp containing an open reading frame (ORF) of 1161bp encoding for a 386 amino acid protein. The full-length cDNA of HpPKS2 was 1559bp with an ORF of 1182bp encoding for a 393 amino acid protein. The highly conserved catalytic amino acid residues common to plant-specific PKSs were preserved in both genes. HpPKS1 and HpPKS2 exhibited distinct tissue-specific expression patterns in H. perforatum. The HpPKS1 expression was highest in flower buds and lowest in root tissues. The expression of HpPKS2 was found to be high in flower buds and leaf margins and low in leaf interior parts, stems and roots. The expression of the HpPKS1 was found to correlate with the concentrations of hyperforin and adhyperforin while the expression of HpPKS2 showed correlation with the concentrations of hypericins and pseudohypericins in H. perforatum tissues.     

Analysis of physiological and miRNA responses to Pi deficiency in alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.)

Key message

The induction of miR399 and miR398 and the inhibition of miR156, miR159, miR160, miR171, miR2111, and miR2643 were observed under Pi deficiency in alfalfa. The miRNA-mediated genes involved in basic metabolic process, root and shoot development, stress response and Pi uptake.

Abstract

Inorganic phosphate (Pi) deficiency is known to be a limiting factor in plant development and growth. However, the underlying miRNAs associated with the Pi deficiency-responsive mechanism in alfalfa are unclear. To elucidate the molecular mechanism at the miRNA level, we constructed four small RNA (sRNA) libraries from the roots and shoots of alfalfa grown under normal or Pi-deficient conditions. In the present study, alfalfa plants showed reductions in biomass, photosynthesis, and Pi content and increases in their root-to-shoot ratio and citric, malic, and succinic acid contents under Pi limitation. Sequencing results identified 47 and 44 differentially expressed miRNAs in the roots and shoots, respectively. Furthermore, 909 potential target genes were predicted, and some targets were validated by RLM-RACE assays. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed prominent enrichment in signal transducer activity, binding and basic metabolic pathways for carbohydrates, fatty acids and amino acids; cellular response to hormone stimulus and response to auxin pathways were also enriched. qPCR results verified that the differentially expressed miRNA profile was consistent with sequencing results, and putative target genes exhibited opposite expression patterns. In this study, the miRNAs associated with the response to Pi limitation in alfalfa were identified. In addition, there was an enrichment of miRNA-targeted genes involved in biological regulatory processes such as basic metabolic pathways, root and shoot development, stress response, Pi transportation and citric acid secretion.

     

SPX proteins regulate Pi homeostasis and signaling in different subcellular level

To cope with low phosphate (Pi) availability, plants have to adjust its gene expression profile to facilitate Pi acquisition and remobilization. Sensing the levels of Pi is essential for reprogramming the gene expression profile to adapt to the fluctuating Pi environment. AtPHR1 in Arabidopsis and OsPHR2 in rice are central regulators of Pi signaling, which regulates the expression of phosphate starvation-induced (PSI) genes by binding to the P1BS elements in the promoter of PSI genes. However, how the Pi level affects the central regulator to regulate the PSI genes have puzzled us for a decade. Recent progress in SPX proteins indicated that the SPX proteins play important role in regulating the activity of central regulator AtPHR1/OsPHR2 in a Pi dependent manner at different subcellular levels.     

草鱼Isthmin-1基因序列分析及转录水平的营养调控

为探讨Isthmin-1(Ism-1)在草鱼糖和脂类代谢中的作用,研究采用RT-PCR技术克隆草鱼Ism-1的开放阅读框(ORF),生物信息学分析Ism-1及其编码的氨基酸序列, RT-qPCR技术检测Ism-1在草鱼各组织中的分布特点,并在细胞和活体水平上分析不同营养条件下Ism-1 mRNA的表达变化。结果显示,成功克隆草鱼Ism-1的ORF区。序列分析表明,草鱼Ism-1基因开放读码框为1380 bp,编码459个氨基酸,预测该蛋白相对分子量为50.96 kD。氨基酸多序列比对和系统进化树分析显示,草鱼Ism-1与黑头软口鲦(Pimephales promelas)的进化关系最近(氨基酸相似度为96.51%)。Ism-1在草鱼各组织中均有表达,在红肌中表达量最高,其次是鳃、脑和白肌等组织。饥饿再投喂实验结果表明,饥饿14d后肝胰脏中Ism-1的表达量显著上调(P<0.05),恢复投喂后表达量降低,但仍显著高于对照组(P<0.05),白肌中Ism-1的表达量在饥饿和再投喂后均显著增加(P<0.05)。腹腔注射不同浓度的胰高血糖素显著下调草鱼肝胰脏中Ism-1的mR...     

德国小蠊两个海藻糖合成酶基因的克隆、组织分布及温度诱导表达分析

【目的】海藻糖合成酶（trehalose-6-phosphate synthase, TPS）是参与昆虫血糖-海藻糖合成的关键酶。本研究旨在克隆德国小蠊 Blattella germanica TPS基因,研究TPS基因在德国小蠊不同组织中的表达模式及在不同温度处理下的表达情况。【方法】通过RACE技术克隆德国小蠊TPS基因全长序列,利用荧光定量PCR的方法检测TPS基因在德国小蠊5龄幼虫不同组织中的表达模式及在高温(40℃和46℃处理30 min)及低温(0℃和10℃处理1 h)逆境下的表达量变化。【结果】从德国小蠊中克隆获得2个TPS基因,分别命名为 BgTPS1 (GenBank登录号：KR050213) 和 BgTPS2 (GenBank登录号：KR050214)。其中,BgTPS1基因cDNA序列全长2 987 bp,开放阅读框 (ORF) 2 502 bp,编码833个氨基酸;BgTPS2基因cDNA序列全长3 212 bp,开放阅读框2 469 bp,编码822个氨基酸。BgTPS1和BgTPS2基因都主要在5龄幼虫脂肪体中表达,且BgTPS2基因的表达量为BgTPS1基因表达量的3.9倍。在两种不同极端温度诱导下,BgTPS1和BgTPS2基因mRNA均上调表达。其中,BgTPS2 的表达量始终显著高于 BgTPS1。在0℃时,BgTPS1和BgTPS2的表达量最高。【结论】德国小蠊5龄幼虫中存在2个TPS基因。两个TPS基因均在脂肪体中高表达,且BgTPS2基因的表达量显著高于BgTPS1基因;低温和高温诱导下均能促进两个基因的表达量上升。该结果为进一步明确昆虫海藻糖的合成途径及其在昆虫对温度逆境的反应中的作用研究奠定了基础。     

Characterization of the promoter of phosphate transporter TaPHT1.2 differentially expressed in wheat varieties

TaPHT1.2 is a functional, root predominantly expressed and low phosphate （Pi） inducible high-affinity Pi transporter in wheat, which is more abundant in the roots of P-efficient wheat genotypes （e.g., Xiaoyan 54） than in P-inefficient genotypes （e.g., Jing 411） under both Pi-deficient and Pi-sufficient conditions. To characterize TaPHT1.2 further, we genetically mapped a TaPHT1.2 transporter, TaPHT1.2-D1, on the long arm of chromosome 4D using a recombinant inbred line population derived from Xiaoyan 54 and Jing 411, and isolated a 1,302 bp fragment of the TaPHT1.2-D1 promoter （PrTaPHT1.2-D1） from Xiaoyan 54. TaPHT1.2-D1 shows collinearity with OsPHT1.2 that has previously been reported to mediate the translocation of Pi from roots to shoots. PrTaPHT1.2-D contains a number of Pi-starvation responsive elements, including P1BS, WRKY-binding W-box, and helix-loop-helix-binding elements. PrTaPHT1.2-D1 was then used to drive expression of 13-glucuronidase （GUS） reporter gene in Arabidopsis through Agrobacterium-mediated transformation. Histochemical analysis of transgenic Arabidopsis plants showed that the reporter gene was specifically induced by Pi-starvation and predominantly expressed in the roots. As there is only one SNP between the TaPHT1.2-D1 promoters of Xiaoyan 54 and Jing 411, and this SNP does not exist within the Pi-starvation responsive elements, the differential expression of TaPHT1.2 in Xiaoyan 54 and Jing 411 may not be caused by this SNP.