Variation of Rice Tungro Viruses: Further Evidence of Two Rice Tungro Bacilliform Virus Strains and Possibly Several Rice Tungro Spherical Virus Variants

Rice tungro disease is caused by two viruses: rice tungro spherical virus (RTSV) and rice tungro bacilhform virus (RTBV). Our results obtained using polymerase chain reaction (for RTBV) and western blot analysis (for RTSV) to study the epidemiology of tungro supported earlier studies that two RTBV strains. South East Asian and Indian, can be differentiated and also better defined the geographic distribution of these two strains.
Data on RTSV variation were not so conclusive and consistent as those on RTBV because of the high degree of microvariation of RNA genomes. Our approach for differentiation of RTSV led to three variants being identified, the geographic distribution of which does not correlate with that found for strains of RTBV.     

Physiology of Rice Tungro Virus Disease: Increased Cytokinin Activity in Tungro-Infected Rice Cultivars

Tungro virus infection decreased the chlorophyll content in the susceptible rice (Oryza sativa) cultivar ‘Taichung Native 1’, while it increased the pigment in the less-susceptible cultivar ‘IR 20’ during the later stages of infection. The virus was recovered from the infected susceptibile cultivar as early as 3 days after inoculation. Relatively low amounts of virus were recovered from the less-susceptible cultivar between 12 and 30 days after inoculation. The senescence of detached leaves of virus-infected less-susceptible cultivar incubated in water in darkness was considerably delayed compared to healthy leaves. The cytokinin activity in the extracts from inoculated less-susceptible cultivar was 69% higher than in the extracts from the healthy plants.     

中国东北地区水稻主要栽培品种的遗传多样性分析

利用68对SSR引物对91份粳稻品种进行了遗传多样性分析。研究结果共检测到293个等位基因,平均4．3个;平均多态信息含量（PIC）为0．313,变动范围为0．022～0．825。RM333和RM206的等住基因数最多,分别为14、10;且PIC也最高,分别为0．825、0．805。聚类和群体差异分析结果表明,东北三省水稻品种的遗传基础狭窄。黑龙江省和吉林省、黑龙江省和日本、吉林省和日本的水稻品种间遗传距离都很小．分别为0．083、0．084、0．090,而辽宁省与吉林省、黑龙江省的水稻品种遗传基础有一些差异。9个地理来源的品种聚类结果,可分为5个大类群,黑龙江省、吉林省、日本和韩国形成第Ⅰ类群;北京和辽宁省归为第Ⅱ类群;中国台湾、云南省、美国分别为第Ⅲ、第Ⅳ和第Ⅴ类群。东北三省是重要的粳稻生产基地,但遗传基础非常狭窄,要克服遗传脆弱性应从地理位置较远的国家或地区收集更丰富的遗传资源。     

Genetic Analysis of Tolerance to Rice Tungro Bacilliform Virus in Rice (Oryza sativa L.) Through Agroinoculation

Balimau Putih [an Indonesian cultivar tolerant to rice tungro bacilliform virus (RTBV)] was crossed with IR64 (RTBV, susceptible variety) to produce the three filial generations F₁, F₂ and F₃. Agroinoculation was used to introduce RTBV into the test plants. RTBV tolerance was based on the RTBV level in plants by analysis of coat protein using enzyme‐linked immunosorbent assay. The level of RTBV in cv. Balimau Putih was significantly lower than that of IR64 and the susceptible control, Taichung Native 1. Mean RTBV levels of the F₁, F₂ and F₃ populations were comparable with one another and with the average of the parents. Results indicate that there was no dominance and an additive gene action may control the expression of tolerance to RTBV. Tolerance based on the level of RTBV coat protein was highly heritable (0.67) as estimated using the mean values of F₃ lines, suggesting that selection for tolerance to RTBV can be performed in the early selfing generations using the technique employed in this study. The RTBV level had a negative correlation with plant height, but positive relationship with disease index value.     

The Differential Reaction of Rice Hybrids to Tungro Virus by Phenotyping and PCR Analysis

Twenty popular rice hybrids were used to screen for rice tungro virus (RTV) disease reaction. Virulent green leafhoppers (GLH) were used as vector to introduce RTV to the rice hybrids. Virus symptoms scores were recorded at 14, 21, 34, 41 and 59 days postinoculation (DPI), which suggested that virus symptoms are greatly influenced by growth stage of plants. To confirm the presence of virus, polymerase chain reaction (PCR)‐based detection of Rice tungro bacilliform virus (RTBV) was carried out at 7, 14, 21 and 59 DPI using virus genome‐specific primers. Virus presence was observed in all the rice hybrids and check varieties, particularly at later stages of infection. This study shows that phenotyping for tungro virus resistance in rice hybrids at 21 DPI gives most reliable results based on both virus symptoms and presence of virus. Further, to assess the relative difference in population of RTBV, quantitative PCR was performed in all the genotypes at 21 DPI. Yield data were also recorded from control and virus‐infected plants to estimate yield loss percentage due to tungro disease. This study is important to understand the response of rice hybrids to tungro virus disease. Results obtained in this study emphasize that molecular detection of virus is very important to screen the rice plants accurately for tungro disease reaction.     

Global Analysis of Rice Tungro Spherical Virus Coat Proteins Reveals New Roles in Evolutionary Consequences

Rice tungro disease is caused by a combination of two viruses: Rice tungro spherical virus (RTSV) and Rice tungro bacilliform virus (RTBV). RTSV has a capsid comprising three coat proteins (CP) species. Three CP genes of RTSV-AP isolate were sequenced and compared with 9 other isolates reported worldwide for their phylogenetic survey of recombination events which revealed that in general Indian isolates are forming one separate cluster while those of Philippines and Malaysia forming a different cluster. A significant proportion of recombination sites were found in the CP1 gene, followed by CP2 and CP3 suggesting that it is a major phenomenon in the evolution of various isolates of RTSV. Some interesting domains and motifs such as; 3,4-dihydroxy-2-butanone 4-phosphate synthase in CP1, Type 1 glutamine amidotransferase domain and RNA binding motifs in CP2, domains of receptor proteins in CP3, and glycosylation motif in CP2 and CP3 were also obtained in RTSV coat protein. In addition, simple modular architecture research tool (SMART) analysis of coat proteins of RTSV predicted the coat protein domain of calicivirus suggesting evolutionary linkages between plant and animal viruses. This study provides an opportunity to establish the molecular evolution and sequence-function relationship of RTSV.     

Ultraviolet Devitalization of Eight Selected Enteric Viruses in Estuarine Water

The effect of ultraviolet (UV) radiation on the devitalization of eight selected enteric viruses suspended in estuarine water was determined. The surviving fractions of each virus were calculated and then plotted against the UV exposure time for purposes of comparison. Analytical assessment of the survival data for each virus consisted of least squares regression analysis for determination of intercepts and slope functions. All data were examined for statistical significance. When the slope function of each virus was compared against the slope function of poliovirus type 1, the analytical findings indicated that poliovirus types 2 and 3, echovirus types 1 and 11, and coxsackievirus A-9 exhibited similar devitalization characteristics in that no statistically significant difference was found (P > 0.05). Conversely, the devitalization characteristics of coxsackievirus B-1 and reovirus type 1 were dissimilar from those of poliovirus type 1 in that a statistically significant difference was found between the slope functions (P < 0.05). This observed difference in devitalization of coxsackievirus B-1 and reovirus type 1 was attributed primarily to the frequency distribution of single and aggregate virions, the geometric configuration, the size of the aggregates, and the severity of aggregation. The devitalization curve of coxsackievirus B-1 was characteristic of a retardant die-away curve. The devitalization curve of reovirus type 1 was characteristic of a multihittype curve. The calculated devitalization half-life values for poliovirus types 1, 2, and 3; echovirus types 1 and 11; coxsackievirus types A-9 and B-1; and reovirus type 1 were 2.8, 3.1, 2.7, 2.8, 3.2, 3.1, 4.0, 4.0 sec, respectively. These basic data should facilitate an operative extrapolation of the findings to the applied situation. It was concluded that UV can be highly effective and provide a reliable safety factor in treating estuarine water.     

Physiology of Rice Tungro Virus Disease: Involvement of Abscisic Acid-Like Substance in Susceptible Host-Virus Interactions

Stunting was severe in susceptible rice (Oryza saliva L.) cultivar ‘Taichung Native 1’ infected with tungro virus (RTV) compared to less-susceptible cultivar ‘IR 20’. The senescence of detached leaves of RTV-infected susceptible cultivar incubated in water in dark was accelerated compared to the healthy leaves as measured by the loss of total chlorophyll content. The transpiration rate of RTV-infected leaves of the susceptible cultivar was much lower than the healthy and RTV-infected leaves of the less-susceptible cultivar. Partially purified extracts obtained from RTV-infected leaves effectively inhibited GA-induced α-amylase synthesis in barley endosperms, and rice seedling growth, and they accelerated senescence of detached rice leaves. In all the three bibassays the ABA-like activity was significantly greater in the extracts from the RTV-infected susceptible cultivar than in extracts from the less-susceptible cultivar.     

Effect of Prehumidification on Sampling of Selected Airborne Viruses

Studies were undertaken to determine if a prewetting device (humidifier bulb) used in combination with an all glass impinger (AGI-30) would increase the recovery of airborne mengovirus-37A, vesicular stomatitis virus (VSV), and the S-13 coliphage. Suspensions of T3 coliphage with mengovirus-37A, VSV, or S-13 were aerosolized and collected by using the AGI-30-humidifier bulb combination to sample the aerosols before and after shifts in relative humidities (RH). These studies revealed the following. (i) At low RH values there was a 3 to 4 log increase in recovery of airborne T3 phage; (ii) concomitantly, the recovery of mengovirus-37A and VSV decreased; and (iii) only at the mid-range RH values was the recovery of S-13 enhanced. The prehumidification technique significantly increased the recovery of airborne T3 phage but decreased the recovery of the two animal viruses tested.     

Cytology and Histology in Somatic Embryogenesis of Indica Rice

The somatic embryogenesis was established from mature dehulled seeds. The histological research showed that embryogenic calli were initiated first from absorbed cells of scutellum of mature seed. And then the embryoids derived from the surface of embryogenic callus. Having been the same structure like a zygotic embryo of rice, the embryoids possessed the major parts of scutellum, coleoptile and coleorhiza. In an embryoid, several developmental stages of pro-embryoid, including single embryogenic cells, two, four and multiple cell stage pro-embryeids and some abnormal embryoids were observed. It could be concluded from this experiment that the embryoid from somatic cell culture in Indica rice possessed an original form of a plant in structure like a zygotic did and derived from a single cell.     

两个水稻品种(系)酵母双杂交cDNA文库的构建和比较分析

"武育粳3号"和"KT95-418"为两个遗传背景基本一致而对水稻条纹叶枯病表现为明显抗性差异的粳稻(Oryza sativa L.ssp. japonica)品种(系).利用SMART技术合成双链cDNA后,通过SfiⅠ酶切位点将cDNA片段定向插入到改造的载体NpGADT7中,构建了两品种(系)水稻的酵母双杂交cDNA文库.检测结果表明:所构建的两个文库库容量均大于1.0×106 cfu;初始文库滴度分别为1.0×1010 cfu/mL和5.0×1010 cfu/mL,扩增文库滴度均为1.0×1011 cfu/mL;两文库重组率均大于95 %;"武育粳3号"文库cDNA插入片段长度集中分布在700 bp～800 bp之间,"KT95-418"集中分布在750 bp～1 000 bp之间;小规模测序结果表明两文库中全长基因的比例均超过60 %.两品种(系)水稻酵母双杂交cDNA文库的构建为筛选分离抗病相关的变异基因及开展寄主与水稻条纹病毒(Rice stripe virus, RSV)互作的研究奠定了基础.     

Occurrence of Aphelenchoides besseyi in Louisiana Rice Seed and Its Interaction with Sclerotium oryzae in Selected Cultivars

Aphelenchoides besseyi, the nematode causal agent of white-tip disease of rice, was recovered from 5.5% of 474 seed samples obtained from rice seed warehouses in Louisiana. Laboratory tests in which A. besseyi-infested rice seed was treated with Phostoxin®, a compound used for control of insects in stored grain, indicate that it also has nematicidal properties. In 18-week-duration greenhouse tests, populations of A. besseyi increased 4-5-fold on the cultivars Saturn and Melrose and 3-fold on Nova ''76. Green weights of Nova ''76 plants inoculated with A. besseyi and Sclerotium oryzae, the causal agent of rice stem rot, were significantly reduced below those of plants inoculated with either organism alone or with distilled water. Weights of Melrose plants were reduced significantly by treatments with A. besseyi alone and A. besseyi plus S. oryzae, but not by S. oryzae alone. Saturn plant weights were not reduced significantly by either organism alone or by the two in combination.     

水稻雄性半不育突变体的细胞学观察

TP-2是由水稻品种台北309自然突变的雄性半不育突变体。解剖小花后观察发现,突变体花药细长,干瘪,白色透明状,雌蕊正常。花粉活力检测结果表明:突变体水稻平均花粉粒活力率为57.599%,1个花粉囊里的花粉粒平均有436粒;正常材料台北309水稻平均花粉活力率为94.177%,1个花粉囊里的花粉粒有798粒。组织切片观察发现:突变体水稻从小孢子母细胞发育到减数分裂结束,和正常株相比较在形态上无显著差异,小孢子形成初期出现异常,绒毡层快速消融,小孢子在其发育过程中因得不到营养不能正常发育,产生的小孢子干瘪,呈不规则状,同时部分小孢子产生破裂消融现象,绒毡层不能正常降解可能是导致TP-2水稻小孢子异常发育的主要原因。通过以上观察,对进一步揭示TP-2突变体的不育机制提供了基本资料,对该材料的组配奠定了基础。