First Report of Rhizoctonia solani AG‐7 on Cotton in Egypt

Eighty‐two isolates of Rhizoctonia solani were recorded from roots of naturally‐infected seedlings of the Egyptian cotton (Gossypium barbadense L.). Anastomosis groups (AGs) of the isolates were determined by using 13 different AGs testers. Three (3.7%) of the isolates were identified as R. solani AG7, while the remaining isolates were belonging to the AG 2‐1, AG4 and AG5. The identification of the three isolates was based on the frequency of the C2 reaction with the AG7 tester isolate. No fusion was observed between AG7 and isolates representing the other 13 AGs. Colonies of AG7 isolates grown on potato dextrose agar (PDA), malt yeast agar (MYA) and melt peptone agar (MPA) were brown to dark brown with aerial mycelium and sclerotia. The isolates had pitted sclerotial clusters and brownish exudates after 21 days of culturing on PDA, but without clear zonation. Pathogenicity test under greenhouse conditions revealed that AG7 caused the common symptoms of damping–off, which included seed rot, lesions on the hypocotyls and root rot.     

Anastomosis Group Determination of Rhizoctonia solani Kühn (Telemorph: Thanatephorus cucumeris) Isolates from Tomatoes Grown in Aydin,Turkey and their Disease Reaction on Various Tomato Cultivars

In the Province of Aydin‐Turkey most frequently Fusarium spp. and secondly Rhizoctonia solani Kühn were isolated from the roots and crown of tomato plants. Based on affinities for hyphal fusion with test isolates, all R. solani isolates were identified as AG‐4. The tomato cultivars which were grown in soil infested with R. solani AG‐4 exhibited different reactions. From the point of symptom expression and the rate of seedling emergence Sunny 6066 F1 was found to be the most resistant cultivar, whereas Rio Grande, Rio Fuego, NDM 725, Interpeel and Konia were the most susceptible cultivars.     

Characterization and Pathogenicity of Rhizoctonia spp. from Onion in Amasya, Turkey

Forty‐two isolates of Rhizoctonia spp. were obtained from onion in Amasya, Turkey. Of these, 29% were Rhizoctonia solani (AG‐4), 69% were Waitea circinata var. zeae (Rhizoctonia zeae) and 2% were binucleate Rhizoctonia (AG‐B). Most of the isolates were recovered from rhizosphere soil. In pathogenicity tests on onion, R. solani AG‐4 caused the greatest disease severity, those of W. circinata var. zeae were moderately virulent but binucleate Rhizoctonia isolates were of low virulence. This is the first report of binucleate Rhizoctonia AG‐B and W. circinata var. zeae occurring on onion in Turkey.     

中国东北地区玉米纹枯病菌的融合群鉴定

自东北三省采集玉米纹枯病标本300余份,分离获得286个丝核菌菌株。融合群测定及5.8S rDNA-ITS区序列分析结果表明,这些菌株分别属于多核丝核菌的AG1-IA、AG1-IB、AG1-IC、AG4-HG-I、AG4-HG-III、AG-5、WAG-Z群及双核丝核菌的AG-Ba群。其中AG1-IA是优势致病群,占分离菌株总数的38.46%,其次是WAG-Z和AG-5群,分别占26.92%及24.83%。AG4-HG-III群菌株是国内首次从罹病玉米植株上分离得到。自各融合群中选取代表性的菌株进行5.8     

Alternative Hosts for Fusarium spp. Causing Crown and Root Rot of Asparagus in Spain

Asparagus crown and root rot caused by Fusarium oxysporum f.sp. asparagi (Foa), F. proliferatum (Fp) and F. solani (Fs) result in early decline and loss of crop production. The role of several crop species on the survival of the Fusarium spp. was investigated. The root symptoms and plant weight of seven crop species were evaluated after inoculation with each of the three Fusarium spp. The number of colony‐forming units of the Fusarium spp. from root tissues was also determined. Garlic was shown to be a symptomatic host for Foa, Fp and Fs; Fs was also pathogenic to onion. Root colonization of garlic, onion, maize, wheat, potato and sunflower suggested that they are reservoirs of Foa, Fp and Fs from asparagus and demonstrated the importance of crop rotation on the development of this asparagus disease.     

Inheritance of a recessive transgene-associated character controlling albinism in transgenic bean (Phaseolus vulgaris L.)

We identified a transgenic line exhibiting albinism during our work to introduce genes through genetic engineering in dry bean (Phaseolus vulgaris). The transgenic mother plant (R0) presented a normal phenotype and generated albino and normal green plants in the first generation (R1). The segregation ratio of the albino character in the R1 and R2 generations fitted the expected ratio for a character controlled by a single recessive gene linked to a foreign gus gene, suggesting that albinism could be a consequence of insertional mutation caused by introduction of the exogenous gene. Analysis by electron microscope revealed that the albino cells possessed no chloroplasts and a greater number of mitochondria when compared to normal green plants. This transgenic bean line may be used in understanding the genetic control of chloroplast genesis, for acquiring additional knowledge of genomic structure or in physiological studies. This is the first described transgene-associated mutant bean plant.     

Common anastomosis and internal transcribed spacer RFLP groupings in binucleate Rhizoctonia isolates representing root endophytes of Pinus sylvestris, Ceratorhiza spp. from orchid mycorrhizas and a phytopathogenic anastomosis group

Binucleate Rhizoctonia endophytes from the roots of nursery-grown Pinus sylvestris (Scots pine) seedlings and the orchid Goodyera repens from Scots pine forests were characterized on the basis of morphological characters, anastomosis group membership and PCR-assisted ribosomal DNA fingerprinting. Common hyphal and colony morphological traits displayed by the Finnish binucleate Rhizoctonia isolates and a range of Canadian orchid root endophytes enabled them to be placed in the anamorphic genus Ceratorhiza . Five main anastomosis groups were identified and included groups that contained different combinations of Scots pine, G. repens and Canadian Ceratorhiza spp. isolates. Two Scots pine root endophytes anastomosed with a phytopathogenic Japanese tester isolate, confirming their membership of anastomosis group I, which is known to include the teleomorphic species Ceratobasidium cornigerum . Hierarchical cluster analysis of RFLPs in the internal transcribed spacer of ribosomal DNA enabled the division of isolates into one of five RFLP groups. The RFLP and anastomosis groupings were closely correlated; isolates within each of four RFLP groups, which shared 100% RFLP identity, anastomosed in the cross-pairing anastomosis group tests. However, all represented different vegetatively compatible populations (clones) because the diagnostic killing reaction, a cellular vegetative incompatibility response, was identified at hyphal fusion junctions. These findings indicate a high degree of intraspecific variation within Ceratobasidium cornigerum , which includes isolates able to enter into either mutualistic or pathogenic root association with susceptible host plants. The common anastomosis group/RFLP groupings identified also strongly support the hypothesis that conifer tree roots can act as large inoculum reservoirs for these orchid endophytes, allowing the development of inter-plant connections, via commonly shared hyphal linkages, in boreal forest ecosystems.     

Inactivation of the nodH gene in Sinorhizobium sp. BR816 enhances symbiosis with Phaseolus vulgaris L

Sulfate modification on Rhizobium Nod factor signaling molecules is not a prerequisite for successful symbiosis with the common bean (Phaseolus vulgaris L.). However, many bean-nodulating rhizobia, including the broad host strain Sinorhizobium sp. BR816, produce sulfated Nod factors. Here, we show that the nodH gene, encoding a sulfotransferase, is responsible for the transfer of sulfate to the Nod factor backbone in Sinorhizobium sp. BR816, as was shown for other rhizobia. Interestingly, inactivation of nodH enables inoculated bean plants to fix significantly more nitrogen under different experimental setups. Our studies show that nodH in the wild-type strain is still expressed during the later stages of symbiosis. This is the first report on enhanced nitrogen fixation by blocking Nod factor sulfation.     

黄淮海地区夏玉米纹枯病菌的融合群鉴定

从黄淮海地区采集玉米纹枯病标样250余份,分离得到176个丝核菌菌株。融合群测定及5.8SrDNA-ITS区序列分析结果表明,这些菌株分别属于多核丝核菌的AG1-IA、AG1-IB、AG4-HG-I、AG-5、WAG-Z融合群及双核丝核菌的AG-A、AG-Ba融合群。其中AG1-IA是优势融合群,占分离菌株总数的64.20%,其次是AG-Ba,占12.50%,再依次分别是WAG-Z(10.23%)、AGI-IB(5.11%)、AG-4-HG-I(3.98%)、AG-5(2.27%)和AG-A(1.70%)。其中AG-Ba融合群是国内首次在玉米上分离得到。从各融合群中选取代表性的菌株进行5.8S rDNA-ITS区序列分析结果表明,隶属不同融合群或亚群的菌株其5.8S rDNA-ITS区序列存在较大的差异,而相同融合群(或亚群)不同菌株之间其序列的一致性可高达97%-100%。     

黄淮海地区夏玉米纹枯病菌的融合群鉴定

One hundred and seventy-six isolates were obtained from corn sheath blight samples in Huanghuai Plain and Haihe Plain (including Shandong, Henan, Hebei Provinces and Northern regions of Jiangsu Province) of China. Anastomosis group identification and 5.8S rDNA-internal transcribed spacer (ITS) sequence analysis showed that the isolates belonged to multinucleate Rhizoctonia AG1-IA, AG1-IB, AG4-HG-I, AG-5 and WAG-Z and binucleate Rhizoctonia AG-A and AG-Ba. Of these, AG-1-IA was the major anastomosis group(AG)（64.20% of total isolates), followed by AG-Ba (12.50%), WAG-Z (10.23%), AG1-IB (5.11%), AG4-HG-I (3.98%), AG-5 (2.27%) and AG-A (1.70%). AG-Ba was isolated for the first time from maize in China. 5.8S rDNA-ITS sequence analysis showed that the isolates could be distinctly separated based on their AG types. The isolates belonging to the same AG (or sub-AG) showed 97%-100% sequence identity.     

Functional specialization of one copy of glutamine phosphoribosyl pyrophosphate amidotransferase in ureide production from symbiotically fixed nitrogen in Phaseolus vulgaris

Purines are essential molecules formed in a highly regulated pathway in all organisms. In tropical legumes, the nitrogen fixed in the nodules is used to generate ureides through the oxidation of de novo synthesized purines. Glutamine phosphoribosyl pyrophosphate amidotransferase (PRAT) catalyses the first committed step of de novo purine synthesis. In Phaseolus vulgaris there are three genes coding for PRAT. The three full‐length sequences, which are intron‐less genes, were cloned, and their expression levels were determined under conditions that affect the synthesis of purines. One of the three genes, PvPRAT3, is highly expressed in nodules and protein amount and enzymatic activity in these tissues correlate with nitrogen fixation activity. Inhibition of PvPRAT3 gene expression by RNAi‐silencing and subsequent metabolomic analysis of the transformed roots shows that PvPRAT3 is essential for the synthesis of ureides in P. vulgaris nodules.     

Effects of high temperature on nodulation and nitrogen fixation by Phaseolus vulgaris L.

Screening of Rhizobium leguminosarum bv. phaseoli strains showed some that were able to nodulate common beans (Phaseolus vulgaris L.) at high temperatures (35 and 38°C/8 h/day). The nodulation ability was not related to the capability to grow or produce melanin-like pigment in culture media at high temperatures. However, nodules formed at high temperatures were ineffective and plants did not accumulate N in shoots. Two thermal shocks of 40°C/8 h/day at flowering time drastically decreased nitrogenase activity and nodule relative efficiency of plants otherwise grown at 28°C. Recovery of nitrogenase activity began only after seven days, when new nodules formed; total incorporation of N in tops did not recover for 2 weeks. Non-inoculated beans receiving mineral N were not affected by the thermal shock, and when growing continuously at 35 or 38°C had total N accumulated in shoots reduced by only 18%.     

Time course of N2 fixation in common bean (Phaseolus vulgaris L.)

Field and greenhouse experiments were conducted to assess the nitrogen fixation rates of four cultivars of common bean (Phaseolus vulgaris L.) at different growth stages. The ¹⁵N isotope dilution technique was used to quantify biological nitrogen fixation. In the greenhouse, cultivars M4403 and Kallmet accumulated 301 and 189 mg N plant^–1, respectively, up to 63 days after planting (DAP) of which 57 and 43% was derived from atmosphere. Under field conditions, cultivars Bayocel and Flor de Mayo RMC accumulated in 77 DAP, 147 and 135 kg N ha^–1, respectively, of which approximately one-half was derived from the atmosphere. The rates of N₂ fixation determined at different growth stages increased as the plants developed, and reached a maximum during the reproductive stage both under field and greenhouse conditions. Differences in translocation of N were observed between the cultivars tested, particularly under field conditions. Thus, the fixed N harvest index was 93 and 60 for cultivars Flor de Mayo and Bayocel, respectively. In early stages of growth, the total content of ureides in the plants correlated with the N fixation rates. The findings reported in the present paper can be used to build a strategy for enhancing biological N₂ fixation in common bean.     

Distribution and impact of virus associated diseases of common bean (Phaseolus vulgaris L.) in northern Iran

Different viral diseases infect common bean crops in Iran. A total of 248 symptomatic samples were collected from common bean fields throughout main growing fields of Guilan province in Iran during the summer of 2006. Eight viruses were detected using double antibody-sandwich – enzyme-linked immunosorbent assay (DAS-ELISA). Bean common mosaic virus – BCMV (1%), Bean leaf roll virus – BLRV (9%), Cowpea mild mottle virus – CpMMV (6%), Southern bean mosaic virus – SBMV (3%), Cucumber mosaic virus – CMV (15%), Bean golden mosaic virus – BGMV (2%), Bean common mosaic necrosis virus – BCMNV (1%) and Bean yellow mosaic virus – BYMV (1%) were detected. Comparatively CMV (15%) was found to be more prevalent in Guilan province. Multiple infections of viruses were recorded in many samples. Weed species belonging to Chenopodiaceae, Solanaceae, Malvaceae and Amaranthaceae families were also found to be infected with the viruses.     

Highly polymorphic microsatellite loci in the rice- and maize-infecting fungal pathogen Rhizoctonia solani anastomosis group 1 IA

Ten polymorphic microsatellite loci were isolated and characterized from the rice- and maize-infecting Basidiomycete fungus Rhizoctonia solani anastomosis group AG-1 IA. All loci were polymorphic in two populations from Louisiana in USA and Venezuela. The total number of alleles per locus ranged from four to eight. All 10 loci were also useful for genotyping soybean-infecting R. solani AG-1 isolates from Brazil and USA. One locus, TC06, amplified across two other AG groups representing different species, showing species-specific repeat length polymorphism. This marker suite will be used to determine the global population structure of this important pathogenic fungus.     

Rhizoctonia solani injuries in oilseed crops in Finland and impacts of different crop management practices on disease incidence and severity

A disease survey in Finnish oilseed Brassica (OSR) fields in 2007–09 revealed the widespread occurrence and several fold increase of necrotic stem base lesions and severely injured blackened roots in comparison to a corresponding survey carried out in 1984–89. Rhizoctonia solani was the predominant fungi detected in the isolations and was followed by several species of Fusarium and Thielaviopsis basicola. In 60% of the samples all three species were detected together. Only the R. solani AG 2–1 strains isolated from OSR and other cruciferous hosts caused damping off or stem base symptoms on turnip rape in a greenhouse experiment. Therefore R. solani AG 2–1 was considered the main pathogen associated with the observed symptoms in OSR crops. Cultural practices changed significantly between the 1980s and 2000s. In the 2007–09 survey there was an increase in the cultivation of oilseed rape instead of turnip rape, increase in the use of no soil or reduced soil tillage and of chemical control of weeds, but a reduction in macronutrient fertilization, especially P and K, when compared to the 1980s survey. The risk for high incidence of stem base lesions and blackened roots was affected by different cultural practices. No tillage and maintaining sufficient soil pH and NPK fertilisation decreased the risk for both types of R. solani induced symptoms. Late sowing date increased the risk for high incidence of stem base lesions, while application of fungicides against Sclerotinia reduced it. The incidence of R. solani damages in many fields was very high in spite of relatively long crop rotations and therefore the average effect of crop rotation in the disease was insignificant. Current turnip rape cultivars are vulnerable to root rot while oilseed rape is vulnerable to stem base symptoms. The higher incidence of R. solani induced diseases could be associated with the decline in productivity of OSR crops in Finland. This study showed that cultural practices such as reduced or no soil tillage, adequate levels of pH and of NPK fertilization could reduce the severity of the symptoms in OSR fields.     

Developmental Regulation of Susceptibility and Tolerance to Fusarium Root Rot in Beans

The soil-borne fungus, Fusarium solani f. sp. phaseoli, attacks roots and hypocotyls of bean (Phaseolus vulgaris) plants causing a devastating disease called root and foot rot. In a study of the host-pathogen relationship it was found that young bean roots, with the radicle just emerging, were highly tolerant to the pathogen, whereas older bean seedlings, with a fully developed root system, were completely susceptible. Investigations by low-temperature scanning electron microscopy demonstrated that significantly fewer spores and hyphae were present on the root surface of young bean seedlings as compared to older ones. A similar pattern of attachment was found when bean roots were inoculated with spores of F. solani f. sp. pisi, a related pathogen causing disease on peas but not on beans. Light microscopic studies showed that F. solani f. sp. pisi did not penetrate the root but rapidly formed thick-walled resting spores on the root surface. F. solani f. sp. phaseoli on the other hand quickly penetrated the root and formed an extensive network of fungal hyphae. These results demonstrate that the ability of fungal propagules to adhere to and to penetrate host tissues are two distinct processes. Furthermore, the data indicate that young bean roots lack a surface component necessary for attachment of fungal spores which may help explain their tolerance to Fusarium root rot.     

Genetics of fertility restoration in cytoplasmic male sterile Phaseolus vulgaris L.

Summary Restoration of fertility in cytoplasmic male sterile Phaseolus vulgaris by line R-351 was controlled by a single gene. The restorer gene (Fr) displayed incomplete dominance leading to partial restoration of fertility in F₁ generations; full restoration was not achieved until the F₂ generation. Once full restoration of fertility was produced in the F₂ generation, no segregation for sterility was observed in subsequent generations derived from heterozygotes Frfr, either by testcrossing (restored × maintainer) or in F₃ progenies. Implications of the irreversible nature of this restoration are discussed.Florida Agr. Exp. Sta. Journal Series No. 7733