桑椹挥发性成分的分析研究

对广东桑桑椹石油醚和乙酸乙酯提取物中的化学成分进行GC-MS分析。与WILEY谱库的标样质谱图进行比较,鉴定出21种化学成分。     

HPLC-ELSD法测定家桑野桑植株中1-脱氧野尻霉素的含量

采用HPLC-ELSD法对桑植株各部位所含的1-脱氧野尻霉素(DNJ)进行了含量测定,使用TSKgel Am-ide-80(4.6 mm×250 mm,5μm)色谱柱,流动相为:乙腈-水(84∶16,6.5 mM醋酸铵),流速为1.00 mL/min,柱温:30℃,线性范围为0.505μg～20.2μg(r=0.9991,n=6),平均回收率为94.95%(n=5),RSD=1.98。测定结果表明,野桑叶、嫩枝和根皮部位的DNJ含量较高,而家桑的叶、枝皮和根皮中含量较高。本方法准确度高、重现性好,能快速检测桑树资源及类似植物和相关产品中1-脱氧野尻霉素的含量。     

Epidemiology and prediction of brown leaf rust of mulberry caused by Peridiopsora mori

Brown leaf rust (BLR) caused by Peridiopsora mori is one of the major foliar diseases of mulberry (Morus sp.) in the subtropical hills of eastern India. The disease appeared in first week of August and continued up to September with maximum severity in second and third week of September. The disease symptoms appeared at atmospheric temperature (27.00–20.07°C), relative humidity (92.14–82.43%), rainfall (11.20 cm) and rainy days (7) of the preceding week. Disease severity (>50 PDI) was observed at temperature (26.29–19.29°C), relative humidity (94.14–80.14%), rainfall (4.12 cm) and number of rainy days (2–3 days). Apparent rate of infection was found high at temperature (27.00–19.83°C), relative humidity (94.67–85.00%), rainfall (4.6 cm) and rainy days (2) of the preceding week. The correlation coefficient between disease severity and average meteorological factors of the preceding 7 days revealed that BLR disease severity showed significant negative correlation with minimum temperature. It was also revealed that contribution of maximum and minimum temperature 42.23% and 35.21%, maximum and minimum relative humidity (RH) 11.23% and 10.69% and rainfall and number of rainy days 0.11% and 0.50%, respectively towards development of BLR disease severity. Multiple regression analysis revealed that average of maximum and minimum temperatures and minimum RH of preceding 7 days were found to maximally influence BLR disease severity.     

Leaf yield loss assessment due to Macrophomina root rot disease in mulberry gardens of south India

Abstract

Leaf yield loss in mulberry due to Macrophomina root rot disease was assessed in three different states of south India at field level. The highest leaf yield loss recorded was in V-1 variety (34.74%), whereas the lowest leaf yield loss was in K-2 variety (28.54%). However, the leaf yields losses in other varieties viz., MR-2 (32.90%), S-36 (32.06%), RFS-175 (31.75%) and S-13 (29.0%) recorded were medium. The average leaf yield loss was 31.49% due to root rot disease caused by M. phaseolina in mulberry.     

Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves

To optimize Agrobacterium-mediated transient transformation assay in mulberry (Morus alba L.), various infiltration methods, Agrobacterium tumefaciens (A. tumefaciens) strains, and bacterial concentrations were tested in mulberry seedlings. Compared with LBA4404, GV3101 harboring pBE2133 plasmids presented stronger GUS signals at 3 days post infiltration using syringe. Recombinant plasmids pBE2133:GFP and pBE2133:GFP:MaFT were successfully constructed. Transient expression of MaFT:GFP protein was found in leaves, petiole (cross section), and shoot apical meristem (SAM) of mulberry according to the GFP signal. Moreover, MaFT:GFP mRNA was also detected in leaves and SAM via RT-PCR and qRT-PCR. An efficient transient transformation system could be achieved in mulberry seedlings by syringe using A. tumefaciens GV3101 at the OD₆₀₀ of 0.5. The movement of MaFT expression from leaves to SAM might trigger the precocious flowering of mulberry.     

Field performance and biochemical evaluation of micropropagated mulberry plants

Microclones of different mulberry genotypes were successfully transferred to the field. The same genotypes were raised through conventional methods (cuttings). A comparative study using morphological and biochemical tests of field established micropropagated and cutting derived plants of mulberry genotypes was conducted. Micropropagated mulberry plants showed significant morphogenic vigour when compared to plants raised through cuttings. Biochemical tests of leaves revealed that, there was no significant nutritional difference between micropropagated plants and those originated from cuttings. This revised version was published online in June 2006 with corrections to the Cover Date.     

桑树内生拮抗菌的分离鉴定及其对桑断枝烂叶病的生防初探

【目的】本研究从健康桑树茎中分离筛选对桑断枝烂叶病菌具显著拮抗作用的内生细菌,为该病生物防治奠定研究基础。【方法】采用组织培养法分离桑树内生菌,抑菌圈法和平板对峙法筛选抑菌活性稳定的内生拮抗菌;根据形态学、生理生化特征检测和基于16SrDNA、gyrA和gyrB基因的系统发育分析对拮抗菌进行菌种鉴定;利用抑菌圈法测定拮抗菌株活性发酵液热稳定性,菌丝生长速率法检测活性发酵液抑菌谱;并通过观察拮抗菌对桑断枝烂叶病菌BoeremiaexiguaGXH1菌株生长及菌丝形态的影响,扩增抑菌活性物质合成关键基因,以及采用酸沉淀法提取拮抗菌株脂肽类化合物并进行高效液相色谱串联质谱分析(LC-MS),初步探究可能的抑菌机制。【结果】从健康桑树茎中共分离获得17株桑树内生细菌,并从中筛选获得一株对桑断枝烂叶病菌B.exiguaGXH1有稳定拮抗作用的桑树内生细菌NPJ13菌株。该菌株形态学、生理生化特征与芽孢杆菌属一致,基于16SrDNA、gyrA和gyrB基因序列的系统发育分析结果显示该菌株与贝莱斯芽孢杆菌(Bacillusvelezensis)的亲缘关系最近,且处于系统发育树的最小分枝,故将NPJ13菌株鉴定为贝莱斯芽孢杆菌,命名为B. velezensis NPJ13。NPJ13菌株对灰霉病菌SWU5、核地杖菌SXSG-5、核盘菌PZ-2及烟草疫霉SWU20等12种病原真菌具有不同程度的拮抗作用,其活性发酵液具有较好的热稳定性。NPJ13菌株会导致桑断枝烂叶病菌GXH1菌丝发生扭曲、膨大、透明度增加、断裂等畸变现象;基因检测结果显示NPJ13菌株基因组中具有PKSI、NRPS、Sfp、ItuD、Srfc等5种抑菌活性物质合成关键基因,LC-MS检测结果表明菌株NPJ13脂肽类粗提物中含有表面活性素和伊枯草菌素。【结论】本研究分离筛选获得一株对桑断枝烂叶病菌具有显著拮抗作用的桑树内生细菌B. velezensis NPJ13菌株,为桑断枝烂叶病的生物防治提供了候选菌株。     

Photosynthetically versatile thin shade leaves: A paradox of irradiance-response curves

Thick sun leaves have a larger construction cost per unit leaf area than thin shade leaves. To re-evaluate the adaptive roles of sun and shade leaves, we compared the photosynthetic benefits relative to the construction cost of the leaves. We drew photosynthetically active radiation (PAR)-response curves using the leaf-mass-based photosynthetic rate to reflect the cost. The dark respiration rates of the sun and shade leaves of mulberry (Morus bombycis Koidzumi) seedlings did not differ significantly. At irradiances below 250 μmol m⁻² s⁻¹, the shade leaves tended to have a significantly larger net photosynthetic rate (P _N) than the sun leaves. At irradiances above 250 μmol m⁻² s⁻¹, the P _N did not differ significantly. The curves indicate that plants with thin shade leaves have a larger daily CO₂ assimilation rate per construction cost than those with thick sun leaves, even in an open habitat. These results are consistently explained by a simple model of PAR extinction in a leaf. We must target factors other than the effective assimilation when we consider the adaptive roles of thick sun leaves.     

Accumulation of pathogenesis-related (PR) 10/Bet v 1 protein homologues in mulberry (Morus bombycis Koidz.) tree during winter

Seasonal evaluation of total soluble protein fractions extracted from cortical parenchyma cells of mulberry (Morus bombycis Koidz.) tree identified a predominant 18 kDa protein that was directly correlated to periods of cold acclimation. The 18 kDa protein, designated as WAP18 (winter accumulating 18 kDa proteins) increased from September to December and then gradually decreased until June. The maximum levels of WAP18 were detected in mid‐winter, which corresponds to the maximum freeze tolerance in cortical parenchyma cells of mulberry tree. Two‐dimensional gel electrophoresis confirmed that WAP18 consists of at least three proteins that range between an isoelectric point of 5.0 and 6.0. All three proteins reacted with anti‐WAP18 antibodies, thereby suggesting that they represent individual isoforms. Furthermore, N‐terminal amino acid sequence analysis demonstrated that all three proteins contain high sequence similarity to each other and high homology to pathogenesis‐related (PR) ?10/Bet v 1 protein families. The purified WAP18 exhibited in vitro cryoprotective activity for the freeze labile l ‐lactate dehydrogenase (LDH) enzyme. These results suggest that WAP18 may function in the freezing tolerance mechanism of cortical parenchyma cells of mulberry tree during winter.