长江上游圆筒吻(鱼句)年龄与生长的研究

于2008—2010年在长江上游四川合江和重庆木洞江段共采集圆筒吻鮈样本511尾,研究了其年龄与生长特征,并提出了合理的鱼类资源保护对策。结果表明,所采集的圆筒吻鮈分6个年龄组,2—4龄居多,占95.7%;体长与鳞径为直线正相关,雌雄群体间无显著差异,表达式为L=51.28 R+37.45(r=0.85,n=397);各龄组、雌雄群体的实测体长均值和退算体长均值无明显差异(t=0.751,P>0.05);体长与体重呈幂函数关系,雌雄群体间无显著差异,表达式为W=8×10 6L3.099(r=0.97,n=397),其生长属等速增长类型;生长规律可以用von Bertalanffy方程表示,分别为:Lt=348.78[1 e 0.18(t+1.15)],Wt=603.17[1 e 0.18(t+1.15)]3.099;生长拐点处的年龄为5.12,此时的体长和体重分别为236.23 mm和180.33 g;圆筒吻鮈个体出现小型化现象;根据研究,长江上游圆筒吻鮈处于不合理的开发状态,应限制捕捞、加强保护。     

溴氰菊酯亚致死浓度对烟草粉螟生物学特性的影响

为探讨溴氰菊酯对烟草粉螟的亚致死效应,本文采用烟叶浸渍法以溴氰菊酯亚致死浓度(LC 10和LC 25)胁迫烟草粉螟3龄幼虫,并通过年龄-龄期两性生命表的方法探究溴氰菊酯胁迫对烟草粉螟发育和繁殖的影响。结果显示:与对照相比,溴氰菊酯LC 10、LC 25处理F 0代的平均单雌产卵量显著下降;LC 25处理F 0代的产卵前期及总产卵前期明显长于对照;LC 25处理与LC 10处理和对照间,F 0代雌、雄虫寿命均差异显著;LC 10处理和LC 25处理F 1代与对照存活率明显下降,F 1代各处理间的卵、1龄幼虫、3龄幼虫、5龄幼虫、7龄幼虫和蛹的发育历期无显著差异;但LC 25处理组2龄、4龄、6龄幼虫历期比对照延长了且差异显著;LC 25处理组的成虫前期比对照组延长了且差异显著。同时LC 25处理的雌、雄虫寿命最短,但3个处理间差异不显著。研究表明,亚致死浓度的溴氰菊酯能够抑制烟草粉螟的生长发育和繁殖,以上研究结果可为田间用药控制烟草粉螟提供理论参考。     

Morphological and molecular identification of Leptosphaeria maculans in canola seeds and flowers collected from the North Iran

Blackleg disease, caused by Leptosphaeria maculans, is one of the most important diseases of rapeseed Brassica napus in Iran as in other regions of the world. The samples including canola petals and seeds were collected during 2014–2015 from canola field in North Iran. Isolates characteristics of fungus were assessed based on the colony growth rate and pycnidia in Potato Dextrose Agar. The pycnidia of the fungus were black, globose to subglobose in shape, the single-celled conidia, hyaline and fusiform with diameters of 4–5 × 1.5–2 μm. Most of the isolates were produced pigment in the liquid culture in variable color brown to black. Thirteen isolates were then separated into pathogenicity groups based on the interactions on B. napus differential cultivars. For the direct detection of seed contamination with L. maculans, PCR was developed using specific primers pair (LmacF, LmacR) which can amplify ITS1 and ITS2 along with the 5.8S rRNA region of L. maculans genome. Based on the followed information and sequence analysis, the fungal isolates from these samples were identified as L. maculans. The findings of this research showed that the disease was aggressive and highly distributed from infested seeds to oilseed rape fields.     

Structure,chemistry, and biological activity of pseudophomins A and B,new cyclic lipodepsipeptides isolated from the biocontrol bacterium Pseudomonas fluorescens

Pseudophomins A and B are cyclic lipodepsipeptides isolated from Pseudomonas fluorescens strain BRG100, a bacterium with potential application for biocontrol of plant pathogens and weeds. Their chemical structures were established by a combination of spectroscopic data, X-ray crystallography, and selective chemical degradation. This unique chemical degradation allowed the unambiguous determination of the absolute configuration of the amino acid residue Leu-1, due to gamma-lactam formation followed by selective cleavage of the adjacent N(8)-C(7) bond. To the best of our knowledge this is the first application of gamma-lactam formation to the determination of absolute configuration of an adjacent amino acid. Pseudophomin B showed higher antifungal activity against the phytopathogens Phoma lingam/Leptosphaeria maculans and Sclerotinia sclerotiorum than pseudophomin A, and is likely to be the main component responsible for the antifungal activity of EtOAc extracts of strain BRG100. By contrast, pseudophomin A showed stronger inhibition of green foxtail (Setaria viridis) root germination than pseudophomin B.     

Crystal structure of the effector AvrLm4–7 of Leptosphaeria maculans reveals insights into its translocation into plant cells and recognition by resistance proteins

The avirulence gene AvrLm4–7 of Leptosphaeria maculans, the causal agent of stem canker in Brassica napus (oilseed rape), confers a dual specificity of recognition by two resistance genes (Rlm4 and Rlm7) and is strongly involved in fungal fitness. In order to elucidate the biological function of AvrLm4–7 and understand the specificity of recognition by Rlm4 and Rlm7, the AvrLm4–7 protein was produced in Pichia pastoris and its crystal structure was determined. It revealed the presence of four disulfide bridges, but no close structural analogs could be identified. A short stretch of amino acids in the C terminus of the protein, (R/N)(Y/F)(R/S)E(F/W), was well‐conserved among AvrLm4–7 homologs. Loss of recognition of AvrLm4–7 by Rlm4 is caused by the mutation of a single glycine to an arginine residue located in a loop of the protein. Loss of recognition by Rlm7 is governed by more complex mutational patterns, including gene loss or drastic modifications of the protein structure. Three point mutations altered residues in the well‐conserved C–terminal motif or close to the glycine involved in Rlm4‐mediated recognition, resulting in the loss of Rlm7‐mediated recognition. Transient expression in Nicotiana benthamiana (tobacco) and particle bombardment experiments on leaves from oilseed rape suggested that AvrLm4–7 interacts with its cognate R proteins inside the plant cell, and can be translocated into plant cells in the absence of the pathogen. Translocation of AvrLm4–7 into oilseed rape leaves is likely to require the (R/N)(Y/F)(R/S)E(F/W) motif as well as an RAWG motif located in a nearby loop that together form a positively charged region.     

Rapid identification of the Leptosphaeria maculans avirulence gene AvrLm2 using an intraspecific comparative genomics approach

Five avirulence genes from Leptosphaeria maculans, the causal agent of blackleg of canola (Brassica napus), have been identified previously through map‐based cloning. In this study, a comparative genomic approach was used to clone the previously mapped AvrLm2. Given the lack of a presence–absence gene polymorphism coincident with the AvrLm2 phenotype, 36 L. maculans isolates were resequenced and analysed for single‐nucleotide polymorphisms (SNPs) in predicted small secreted protein‐encoding genes present within the map interval. Three SNPs coincident with the AvrLm2 phenotype were identified within LmCys1, previously identified as a putative effector‐coding gene. Complementation of a virulent isolate with LmCys1, as the candidate AvrLm2 allele, restored the avirulent phenotype on Rlm2‐containing B. napus lines. AvrLm2 encodes a small cysteine‐rich protein with low similarity to other proteins in the public databases. Unlike other avirulence genes, AvrLm2 resides in a small GC island within an AT‐rich isochore of the genome, and was never found to be deleted completely in virulent isolates.     

Genetic Relationship of Brassicaceae Hybrids with Various Resistance to Blackleg Is Disclosed by the Use of Molecular Markers

Brassica napus is an important oil source. Its narrow gene pool can be widened by interspecific hybridization with the Brassicaceae species. One of the agronomically important traits, that can be transferred through the hybridization, is the resistance to blackleg, a dangerous disease mainly caused by Leptosphaeria maculans. Hybrid individuals can be analyzed with various molecular markers, including Simple Sequence Repeats (SSR). We investigated the genetic similarity of 32 Brassicaceae hybrids and 19 parental components using SSR markers to reveal their genetic relationship. Furthermore, we compared the field resistance to blackleg of the interspecific progenies. The tested set of 15 SSR markers proved to be useful in revealing the genetic distances in the Brassicaceae hybrids and species. However, genetic similarity of the studied hybrids could not be correlated with the level of field resistance to L. maculans. Moreover, our studies confirmed the usefulness of the Brassicaceae hybrids in terms of blackleg management.     

Morphological Studies of Rhaphidopteris hsüi sp. nov

The present paper deals with the cuticular structure of Rhaphidopteris hsüi sp. nov. The specimens were collected from the Upper Triassic cos series of Liuzhi district, Guizhou Province. Based on the shape and cuticular structure of leaf and segments, this new species is assigned to Corystospermaceae of Cycadofilicales. According to the assemblage of the fossil plants, the writers consider that the geological age of this flora is assigned to the middle Keuper-Rhaetic stage of Late Triassic.     

Changes in protein kinase and protein phosphatase properties during the cycle of asparaginase activity in Leptosphaeria michotti

Regulation of the cyclic activity of asparaginase (obtained as a purified protein complex) by a reversible auto-phosphorylation process has been previously reported in the fungus Leptosphaeria michotii (West) Sacc. In the present study, the protein complex was purified in the presence of either a mixture of 3 protein phosphatase inhibitors (fluoride, vanadate and molybdate) or EGTA, during the cycle of asparaginase activity, and the protein kinase and protein phosphatase activities characterized. (I) At the phase of increasing asparaginase activity, a Ca²⁺/calmodulin-dependent kinase activity was identified by (a) its inhibition by calmidazolium, reversed by calmodulin, and its inhibition by EGTA, but not by poly(Glu/Tyr 4:1)n. dichloro-(ribofuranosyl)-benzimidazole or polylysine (b) an increasing level of calmodulin bound to the complex, as estimated by enzyme-linked immunosorbent assay (ELISA). (2) At the phase of decreasing asparaginase activity, the Ca²⁺-calmodulin-dependent kinase activity disappeared and a little calmodulin remained associated with the complex: phosphorylation of the complex was increased several-fold by 1 nM okadaic acid and 25 nM inhibitor-2, and was not affected by EGTA, indicating a protein phosphatase-2A-like activity. (3) When asparaginase activity was low, a little calmodulin was bound to the complex. The kinase could phosphorylate casein and phosvitin. was inhibited by poly(Glu/Tyr 4:1)n. dichloro-(ribofuranosyl)-benzimidazole and heparin, stimulated by polylysine and not affected by calmidazolium or EGTA, just as a casein kinase 2. A Ca²⁺-dependent but calmodulin-independent protein phosphatase activity, not affected by okadaic acid and inhibitor-2. was then identified. We postulate the presence in the complex, of (a) only one protein kinase and one protein phosphatase, whose properties could change during the cycle of asparaginase activity: (b) two Ca²⁺/-binding proteins: first calmodulin, which could bind to Ca²⁺ and the casein kinase-2 form to give a Ca²⁺/calmodulin-dependent kinase, which could become Ca²⁺/calmodulin-independent following an auto-phosphorylation process: second a protein homologous to calmodulin, able to bind to the protein phosphatase-2A catalytic subunit to give a protein phosphatase-2B catalytic subunit.