Non-abscised aborted sweet cherry fruits are vulnerable to fruit decaying fungi and may be sources of infection for healthy fruits

During three growing seasons (1999–2001), disease incidence of non-abscised aborted and normally developing (sound) sweet cherry fruits were investigated in a research orchard at Ullensvang, western Norway. To reveal possible fungal infections, aborted and sound fruits of two cultivars (cv. Van for three years and cv. Lapins for two years) were harvested over four to seven weeks in an experimental orchard during the green fruit phase and incubated at 20°C in moisture saturated air for 7 days. The most frequently observed fungal pathogens were Monilinia laxa, Colletotrichum gloeosporioides, and Botrytis cinerea. The mean of all observations (±standard deviation) over three years, showed that aborted and sound fruits had a disease incidence of 51.9% (±31.8) and 5.2% (±9.7), respectively, after seven days incubation. In 24 of 25 trials aborted fruits had significantly higher disease incidence than sound fruits after incubation. In one season, when fruits were collected from two commercial orchards, aborted fruits had much higher disease incidence than sound fruits (a mean of 6.5 and 4.5 times higher incidence for the two orchards, respectively). Time of fruit abortion varied with the years (two years observation) and the two cultivars, but the major abortion took place between the fourth and eighth week after anthesis. A higher disease incidence and more rapid disease development in non-abscised aborted fruits indicate that they are more vulnerable to fungal colonisation than sound fruits and may thus be potential incoulum sources for neighbouring, healthy fruits.     

甜樱桃矮化砧木矮化机理解剖学研究

本研究以六种甜樱桃砧木为试材，对其根皮率、根导管面积、根导管密度、根导管总面积／木质部面积比值、根木质部面积／横断面积比值、枝皮率、枝导管面积、枝导管平均总面积／木质部平均面积比值和枝木质部面积／横断面积比值等解剖结构进行了比较。试验结果表明不同砧木间各测定值存在较大差异，可以认为根皮率，枝导管密度、根、枝导管总面积／木质部面积比值和根、枝木质部面积／横断面面积比值可以作为预测甜樱桃矮化砧木矮化性的综合指标，为今后甜樱桃矮化砧木的选育提供依据。     

根际促生细菌混合接种对甜樱桃/东北山樱根际微生物区系及根系呼吸的影响

为探讨有益微生物对甜樱桃根系的接种效应,试验采用灌根的方法,研究了从野生东北山樱根际分离筛选出的4株促生细菌,混合接种对甜樱桃/东北山樱根际微生物区系及根系呼吸代谢途径的影响.结果表明:接种促生菌液改变了甜樱桃根际细菌、放线菌及真菌的数量比例,使根际细菌数量显著增加.接种2次,根际细菌增加幅度最大,为对照的3.4倍;接种促生菌液提高了根系活力,接种2次效果最显著,为对照的2.4倍,达到200.4μg/(g·h);接种促生菌液对根系呼吸的基础生化途径与末端氧化电子传递途径运行比例无明显影响,但整体上提高了根系呼吸速率.     

山东地区樱桃绿环斑驳病毒(CGRMV)的RT-PCR检测及外壳蛋白基因的克隆

为调查我省甜樱桃感染樱桃绿环斑驳病毒(Cherry Green Ring Mottle Virus,CGRMV)情况,本研究以甜樱桃(Prunus avium L.)品种"红灯"叶片总RNA为模板,根据CGRMV基因组序列设计特异引物,对山东地区37份甜樱桃"红灯"样品进行RT-PCR检测,共检测出19份阳性样品。利用CGRMV外壳蛋白基因序列引物,从阳性植物样本中分离到约800 bp的目的片段,克隆测序,序列分析显示该片段全长807 bp,编码268个氨基酸,与GenBank中已登录的CGRMV分离物的外壳蛋白基因序列一致性为87%～97%,氨基酸序列相似性为95%～99%。该结果表明山东地区甜樱桃生产园中感染CGRMV的病例较为普遍。     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.     

DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.     

几种木本园艺植物塑料袋内密封扦插试验

本文报道在食品保鲜袋内密封扦插桂花、樱花、甜樱桃和苹果的试验结果。当年生半木质化至完全木质化插条经生根促进剂处理后，密封扦插在食品保鲜袋内的湿润基质上，放置空旷处，夏秋高温季节晴天适度遮阴。结果表明，塑料袋内的温度较稳定，基质始终保持刚扦插时的湿润状态，插条叶片全部或部分保持饱满、挺立。插条一周左右即可产生愈伤组织，３─４周便可出根。尽管在几种材料上获得的生根率差异较大，但都成功地生了根。在以“峨嵋仙土”为扦插基质，６月份晴天１０：００─１７：００时遮阴的条件下，樱花的生根率达７０％以上。     

Improved discrimination of self-incompatibility S-RNase alleles in cherry and high throughput genotyping by automated sizing of first intron polymerase chain reaction products

A novel polymerase chain reaction (PCR) approach to determine and confirm the self‐incompatibility (S) genotype of cherries is reported. The method involves PCR amplification with a new pair of consensus primers that immediately flank the first intron of cherry S‐RNases, one of which is fluorescently labelled. Fluorescent amplification products range from 234 to c. 460 bp and can be sized accurately on an automated sequencer. Thirteen S alleles reported in sweet cherry can be distinguished, except for S₂ and S₇, which have an amplification product of exactly the same size. S₁₃, which is also amplified, gives a microsatellite‐like trace which shows minor intra‐allelic length variation. This method gives fast and accurate results and should be especially useful for medium/high‐throughput genotyping of wild and cultivated cherries.