Clonostachys rosea,a potential biological control agent for Rhizoctonia solani AG-3 causing black scurf on potato

Rhizoctonia solani AG-3 forms sclerotia on potato tubers, reducing quality and marketability. Potato discs with sclerotia were exposed to saturated soil, and following toothpick baiting, hyphae of R. solani parasitised with mycelia that either coiled around or penetrated cell walls were observed. Similar parasitic behaviour was observed in dual cultures. Clonostachys rosea was identified as the mycoparasite by morphology and ITS sequencing. When co-inoculated onto stems grown from disease-free seed potatoes, tubers of the C. rosea?+?R. solani inoculated plants had significantly lower black scurf (P?0.0001) and higher yield (P?=?0.0002) than R. solani inoculated controls.     

Highly polymorphic in silico-derived microsatellite loci in the potato-infecting fungal pathogen Rhizoctonia solani anastomosis group 3 from the Colombian Andes

Fourteen polymorphic microsatellite DNA markers derived from the draft genome sequence of Rhizoctonia solani anastomosis group 3 (AG-3), strain Rhs 1AP, were designed and characterized from the potato-infecting soil fungus R. solani AG-3. All loci were polymorphic in two field populations collected from Solanum tuberosum and S. phureja in the Colombian Andes. The total number of alleles per locus ranged from two to seven, while gene diversity (expected heterozygosity) varied from 0.11 to 0.81. Considering the variable levels of genetic diversity observed, these markers should be useful for population genetic analyses of this important dikaryotic fungal pathogen on a global scale.     

Hydrolytic Activities of Fusarium solani and Fusarium solani f. sp. eumartii Associated with the Infection Process of Potato Tubers

The development of dry rot caused by Fusarium solani f. sp. eumartii was evaluated in susceptible (Huinkul) and resistant (Spunta) potato cultivars. Fungal proteolytic and polygalacturanase activities were measured at different days postinoculation either with the pathogenic F. solani f. sp. eumartii, isolate 3122 or with the non‐pathogenic F. solani, isolate 1042. After inoculation with the pathogenic fungus, proteolytic and polygalaturonase activities were higher in the susceptible than in the resistant cultivar. In addition, we found a correlation between the levels of proteolytic activity detected in the intercellular washing fluids with the size of the lesion area caused by F. solani f. sp. eumartii in Huinkul tubers. The action of the proteolytic activity over cell wall proteins of both potato cultivars was assayed. An extracellular potato protein with homology to proteinase inhibitors of the Kunitz family was identified as a substrate of the proteolytic activity in the susceptible cultivar. A microscopic study revealed differences between the potato genotypes in the rate of response to infection by F. solani f. sp. eumartii. In addition, the cell wall alteration caused by F. solani f. sp. eumartii in cortical cells of susceptible tubers was evaluated. The data with respect to the correlation between the course of cyto‐ and biochemical events of the two host–pathogen interactions were discussed.     

中国北方马铃薯黑痣病立枯丝核菌的融合群鉴定

从山东、甘肃、青海、内蒙古、河北和黑龙江6省采集马铃薯黑痣病标本300余份,分离获得251个立枯丝核菌Rhizoctonia solani菌株。融合群测定结果表明,这些菌株分别属于多核的丝核菌AG‐3、AG1‐IB、AG4‐HG‐Ⅰ、AG4‐HG‐Ⅱ、AG4‐HG‐Ⅲ、AG‐5和AG‐11融合群。其中AG‐3是优势致病群,占分离菌株总数的71.31%;其次是AG4‐HG‐Ⅰ,占15.14%;AG‐11融合群菌株是国内首次从罹病马铃薯植株上分离得到。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菌株的5.8S rDNA‐ITS区序列存在较大的差异,而相同融合群(亚群)不同菌株的序列具有较高一致性。     

Survival of Sclerotia of Rhizoctonia solani AG3PT and Effect of Soil‐Borne Inoculum Density on Disease Development on Potato

Sclerotia produced by a single isolate of Rhizoctonia solani AG3PT were buried in small plot experiments to investigate the effects of sclerotial production method, soil type and burial depth on sclerotial viability in field soil. The factor with the greatest effect on sclerotial viability, defined as the percentage of sclerotia germinating on agar following retrieval, in all experiments was the duration of burial. After 18 months, on average across all experiments, 20% of retrieved sclerotia were viable. A comparison between sclerotia produced in vitro on malt yeast extract agar and in vivo using micropropagated tubers in field soil found no significant differences between the two production methods on sclerotial viability. Burial in field soil at 20‐cm depth was found to significantly reduce sclerotial viability to 50% compared to 60% at 5 cm. In two pot experiments, amending the growing medium and soil with increasing inoculum densities of R. solani was found to increase stem number, stem canker and black scurf severity regardless of whether this soil‐borne inoculum was derived from mycelium or sclerotia. Black scurf incidence and severity were assessed 30–32 days posthaulm destruction and found to be similar for a range of sclerotial soil‐borne inoculum densities (1.0 × 10^?1 g/kg d.w. soil to 6 × 10^?3 g/kg d.w. soil). The significance of these findings in relation to pathogen survival, detection in soil and disease development is discussed.     

马铃薯不同组织的诱导分化及其对遗传转化效率的影响

在农业快速发展过程中,基因工程作为改造马铃薯性状的重要手段,一直备受关注。优化马铃薯组织培养体系及外源基因转化条件是进行马铃薯转基因工作的基础。以三种熟性不同的马铃薯栽培品种（东农303、早大白、大西洋）的试管苗为试验材料,采用正交实验,对茎段和试管薯的分化体系进行筛选、优化,建立不同品种茎段的愈伤再生体系及试管薯直接分化再生体系。将以LYCB为目的基因,以NPTII为筛选标记的载体,利用农杆菌转化法,分别对三种材料试管苗的茎段及试管薯进行转基因操作,并对转化条件进行优化,以建立适合于不同品种马铃薯茎段及试管薯的遗传转化体系。以研究中得到的最佳离体再生体系及最优遗传转化体系为基础,利用PCR方法,进行阳性植株检测并统计。研究发现,在东农303、早大白、大西洋三种品种中,经由茎段愈伤组织转化,获得的阳性植株转化率分别为36%、35%、28%;经由试管薯直接分化,获得的阳性植株转化率分别为43%、45%、17%。在后期转基因操作中,东农303/早大白转化时可采用试管薯作为试验组织,大西洋则适合用茎段。     

马铃薯人工种子的研究

本研究以虎头（Ｈ）、克四（Ｋ）和Ｆａｖｏｒｉｔａ（Ｆ）三个品种的马铃薯不定芽在液体培养条件下诱导生成的微型薯为试材,经筛选后获得的大小一致的微型薯经过适度的低温处理后,用４％的海藻酸钠和２％的氯化钙溶液,在附加一定的植物激素条件下,进行人。种皮包埋形成人工种子。在无菌条件下,三个品种的人工种子萌发率均达９０％以上。在土壤中,人工种子萌发率可达６０％以上,萌发的人工种子中８０％能够形成生长发育正常的完整植株。     

Changes in the Activity of Polygalacturonase-Inhibiting Protein during Potato Development

The activity of a polygalacturonase-inhibiting protein was determined in growing potato plants and in stored potato tubers. The activity in leaves was higher than in stems, and it decreased by the end of the vegetative season. During the dormancy period, the inhibitory activity in tubers also changed. In the sprouting tubers, it was somewhat lower than in the nonsprouting ones, and, in sprouts, it was usually higher than in tubers. Both the plant polygalacturonase and the polygalacturonase secreted by phytopathogenic fungi after their penetration in plant tissues can serve as inhibitor's targets. Therefore, the inhibitor seems to control the resistance of plants to infection by particular pathogens, and this resistance is characteristic of definite developmental stages.     

Effect of Potassium on Potato Microtuber Production in vitro

The role of potassium nutrition in microtuber production was investigated in two potato cultivars belonging to different maturity groups. Alterations were made in Murashige and Skoog (MS) medium to obtain seven concentrations of potassium (10, 15, 20, 25, 30, 35 and 40 mM), and their effects were studied on microtuber number, microtuber mass and harvest index. In late cultivar Kufri Sindhuri, an increase in potassium concentration beyond 25 mM decreased the number of microtubers. Whereas, potassium did not show any inhibitory effect on microtuber number in early cultivar Kufri Ashoka. Potassium showed promoting effect on microtuber mass in both the cultivars. Maximum microtuber mass and harvest index were observed when the medium was supplemented with 40 mM potassium. Therefore, large size potato microtubers amenable to direct field planting can be induced in MS medium containing 40 mM potassium. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Influence of Pratylenchus penetrans on the Incidence and Severity of Verticillium Wilt of Potato

The influence of Pratylenchus penetrans on the incidence and severity of Verticillium wilt was examined in the potato cultivars ''Kennebec'', ''Katahdin'', and ''Abnaki''. Single-stem plants were grown in soil maintained at a temperature of 22 ± 1 C. Axenically cultured nematodes were suspended in water and introduced to the soil, at a rate of ca 5,000/25.4-cm pot, through holes made around each stem. Ten days after infestation with nematodes, conidial suspensions of Verticillium albo-atrum were introduced into the soil at a rate of ca 1,000,000/pot. Among Katahdin plants, the severity of foliar symptoms was increased in the presence of both pathogens 2 and 3 weeks after soil intestation. During the remaining 5 weeks, severity of foliar symptoms was not different between plants infected by both pathogens and those infected by Verticillium alone. Within the wilt-susceptible cultivar Kennebec and the resistant eultivar Abnaki, no effects on foliar symptom severity were observed. When plant heights, shoot weights, and tuber yields were analyzed, a Pratylenchus-Verticillium interaction was not evident within any of the cultivars tested. Nematode populations in roots and rhizosphere were suppressed in Kennebec and Katahdin plants in the presence of Verticillium.     

Biology,Epidemiology and Management of Rhizoctonia solani on Potato

Black scurf and stem canker on potato is an economically important disease complex, causing both quantitative and qualitative damage to potato crops which occurs in potato production areas throughout the world. The ribosomal DNA internal transcribed spacer sequence analysis is currently accepted and a commonly used method for classifying Rhizoctonia species and anastomosis groups (AGs). To date, 13 AGs have been recognized. The updated AG distribution in potato worldwide production areas confirm the status of AG‐3 as the most prevalent AG in potato and reflects the population dynamics of the pathogen probably due to global trading of tubers. As R. solani is a tuber‐ and soilborne pathogen, the ability to detect its levels in the seed tubers and in the soil and predict the potential damage is an important factor in controlling the disease. Effective disease management of Rhizoctonia disease requires implementation of an integrated disease management approach and knowledge of each of its stages. Although the most important control measures are cultural, chemical control (either by seed tuber‐ or in‐furrow treatments) is still an important tool in reducing the damages caused by R. solani.     

Influence of osmotic stress on the respiration of potato tuber callus

Potato tuber ( Solanum tuberosum L. cv. Bintje) callus shows a decrease in fresh weight and an increase in dry weight upon transfer to nutrient medium supplemented with 0.3 or 0.5 M mannitol. The osmolarity of the intracellular fluid increases simultaneously. Probably mannitol is taken up from the medium till the osmolarity of the tissue is in equilibrium with that of the medium. After osmotic adaptation, on a medium with 0.5 M mannitol, growth is negligible, although the tissue retains its viability.
Respiration increases upon transfer to medium with extra mannitol, especially when expressed on a fresh weight basis. On this basis cytochrome and alternative pathway capacities do not change appreciably. The respiratory increase is exclusively caused by an increased engagement of the alternative pathway. The participation of this pathway in uninhibited respiration increases from about 10 to 90% upon transfer to medium with extra mannitol. The increase in respiration is partly correlated with the decrease in fresh weight upon transfer. Per disc, the capacities of the cytochrome and alternative pathway decline. Yet, total respiration per disc significantly increases due to the increased participation of the alternative pathway. This results in an almost equal ATP-production per disc before and after transfer. We suggest, that the alternative pathway functions as a reserve capacity in potato callus, which is switched on when ATP-production coupled to the cytochrome pathway is impaired.     

Activity of Membrane-Associated Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase during Development of the Inner Membrane System in Potato Chloroplasts

Membranes of etioplasts and chloroplasts from potato plants (Solanum tuberosumL., cv. Zhu-kovskii) were fractionated by centrifugation in a sucrose gradient, and the fractions of primary membranes, fretlike structures, and frets and granal thylakoids were isolated. The carboxylase and oxygenase activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were measured in these fractions. The membrane-bound Rubisco (mRubisco) was reconstructed in the artificial system from the primary membranes and the soluble form of Rubisco (sRubisco), both isolated from the chloroplasts. The higher carboxylase and lower oxygenase activities were found to be characteristic of this reconstructed mRubisco as compared to sRubisco. The degree of sRubisco association with primary membranes depended on the Mg²⁺concentration. The data suggested that the association of sRubisco with primary membranes occurred at the early stages of membrane formation. The biological role of mRubisco is assumed to consist in controlling the ratio between photosynthesis and photorespiration at the membrane level in plants of various physiological states.     

不同生理年龄马铃薯种薯芽中的内源激素含量变化及其对马铃薯植株生长发育的影响

测定云南马铃薯主栽品种‘合作88’不同生理年龄种薯芽中内源激素含量的结果表明,随着马铃薯种薯生理年龄的增加,各种内源激素含量均呈下降趋势,生理年龄30d以后的种薯中,IAA、GA_3,玉米素(ZR)和双氢玉米素(DHZR)下降显著,ABA下降相对缓慢;生理年龄30d的种薯,主茎长出的叶片数多,单株薯重高。     

水杨酸对陈化马铃薯切片交替途径活性的诱导作用

在马铃薯切片陈化过程中,总呼吸速率（Ｖｔ）和交替途径容量（Ｖａｌｔ）分别升高了５倍和２８倍,但交替途径在无ＫＣＮ存在下几乎不运行。用２０μｍｏｌ／Ｌ的水杨酸处理陈化切片可引起总呼吸速率和交替途径容量的少量增加,但交替途径的运行却明显加强。水杨酸的这种促进作用在Ｐｅｒｃｏｌｌ纯化的线粒体中也有出现。马铃薯切片中交替途径运行的增加导致了线粒体呼吸控制比及ＡＤＰ／Ｏ的下降。这些都表明水杨酸对马铃薯切片陈化过程中抗氰交替途径的活性具有诱导作用。     

Tuber Formation and Growth of in vitro Cultivated Transgenic Potato Plants Overproducing Phytochrome B

We studied the effect of the ectopic expression of the Arabidopsis PHYB gene, which encodes the phytochrome B (phyB) apoprotein, under the control of cauliflower mosaic virus 35S promoter on the photoperiodic response of tuberization and growth of potato (Solanum tuberosum L., cv. Désirée) transformed lines. Stem cuttings of transformed and control plants were cultured on Murashige and Skoog nutrient medium containing 5 or 8% sucrose in the phytotron chambers at 20°C under conditions of a long day (16 h), a short day (10 h), or in darkness. We showed that the overexpression of the PHYB gene enhanced the inhibitory effect of the long day on tuberization. In addition, tuber initiation in these transformed plants occurred at a higher sucrose concentration. The insertion of the PHYB gene decreased plant and tuber weights and shortened stems and internodes. Thus, we demonstrated the complex result of the PHYB gene insertion: it affected the photoperiodic response of tuberization, the control of tuber initiation by sucrose, and the growth of potato vegetative organs.     

Effect of Zinc Sulfate and Boric Acid on the Hormonal Status of Potato Plants in Relation to Tuberization

Potato (Solanum tuberosum L.) plants were grown in a greenhouse using zinc- and boron-deficient soil. The effects of seed-tuber treatment with 3 mM zinc sulfate and 8 mM boric acid on the content and ratio of phytohormones in the leaves and mature tubers, the indices of photosynthetic activity, the rate and NaF-sensitivity of respiration, and the tuber growth were studied. Zinc-sulfate treatment shifted the hormonal balance toward a substantial increase in the cytokinin content and the cytokinin/ABA ratio, as well as a decrease in the IAA/cytokinin ratio. Boric-acid treatment resulted in an increase in the IAA content and IAA/cytokinin ratio. Zinc-sulfate treatment abolished the apical dominance and increased the tuber weight due to their increased number and the number of phellem (cork) cell layers. Boric-acid treatment increased cell diameter in the tuber perimedullary zone; an increase in tuber weight per plant was related to tuber growth. A relationship between changes in the plant hormonal status induced by zinc-sulfate and boric-acid treatments and the activity of physiological processes is discussed.