Inheritance of a pectate lyase mediated soft rot resistance in crosses between transgenic potato lines of cv. désirée andS. tuberosum cultivars

Summary Expression of the pectate lyase (PL) isoenzyme 3 in transgenic potato lines of cv. Désirée mediates an enhanced resistance of tuber tissue toErwinia carotovora (Ec) soft rot due to a pre-activation of plant defence mechanisms. Therefore, theSolanum tuberosum cvs Agave and Adretta with a moderate level of soft rot resistance were crossed with such PL-expressing potato lines. The resulting progenies were assessed with respect to plant/tuber characteristics over a period of four years and then tested for PL3-expression as well as for soft rot resistance. 71% of the selected progeny lines exhibited a stable production of the PL3 enzyme. Statistical analysis revealed differences between the transgenic and the non-transgenic progeny concerning the soft rot resistance of tuber tissue. Compared with the PL-inactive progeny, extension of Ec-rotting on the wound surface of PL-transgenic potatoes was diminished on average by 51.8%. Similarly, the degree of cell lysis caused by bacterial maceration was significantly (P<0.05) reduced in tuber tissue of PL-expressing progeny lines. The latter also revealed an enhanced PPO and PAL activity in their tuber tissue indicating an active plant defence. It is concluded therefore that the PL-mediated soft rot resistance introduced into potatoes by means of molecular techniques is heritable.     

实时定量荧光PCR法检测马铃薯黑胫病菌

马铃薯黑胫病是国内外马铃薯产区发生比较普遍的一种细菌性病害,严重地影响了马铃薯的产量和质量。本研究根据Potato Black Leg序列,设计出马铃薯黑胫病菌特异性的引物,对10种供试菌株进行了实时荧光PCR检测。结果表明,该方法的特异性好,可以将马铃薯黑胫病菌和其它马铃薯常见病害相区分;灵敏度较高,可检测出最低的黑胫病菌浓度为3.6~3.9 cfu.mL-1;而且检测时间仅用4 h,大大缩短了检测周期。该方法可有效地应用于马铃薯黑胫病菌的检测和监控。     

利用体细胞杂交获取马铃薯软腐病的抗性

对马铃薯四倍体栽培种和二倍体野生种Solanumbrevidens的体细胞杂种通过叶片离体培养获得的四倍体植株,以及用四倍体栽培种进行回交获得的五倍体植株的块茎对软腐病的抗性进行了测定。结果表明,由体细胞杂种通过叶片组织离体培养再生植株中,有一个株系SC107对软腐病菌Erwiniacarotovora具有较强的抗性。在用不抗软腐病的马铃薯栽培种对体细胞杂种进行回交获得的杂种后代中,大部分株系对软腐病菌具有高水平的抗性,从而说明Solanumbrevidens对软腐病的抗性基因已转移到马铃薯栽培种。     

Transgenic potatoes expressing anErwinia pectate lyase gene — results of a 4-year field experiment

Summary Four potato lines of cv. Désirée that express the pectate lyase (PL) isoenzyme 3 ofErwinia carotovora subsp.atroseptica were examined in a 4-year field experiment with respect to plant development, tuber yield and resistance of tuber tissue toErwinia soft rot. The PL3 degrades plant cell wall pectin into unsaturated oligogalacturonates eliciting plant defence responses. In one line, enzyme expression was controlled by the CaMV 35S promoter (C) and in three lines it was driven by the potato patatin B33 promoter (D). Plant development of the D-lines in field plots was not distinguishable from that of the non-transgenic counterpart. Also tuber yield was not too different. By contrast, plants of the C-line were smaller than those of the nontransformed counterpart and also showed reduced tuber yield. There were no significant differences in dry mass, starch and protein content of tuber tissue between PL transgenic and non-transgenic potatoes. But compared with the latter, field-grown tubers expressing the PL displayed an enhanced resistance toErwinia soft rot. Thus, average rotting caused byEc-bacteria was diminished in tubers of PL-transgenic lines by 34.1%. The resistance of tubers toEc soft rot was significantly correlated with the PPO activity in tuber tissue.     

Evaluation of the reliability of determining soft rot resistance in potatoes by the tuber slice method

Soft rot resistance in potato (Solanum tuberosum L.) tubers can be determined by inoculating tuber slices with soft rotErwinia species. Questions have been raised in the literature concerning the reliability of the tuber slice method. The objectives of this study were to 1) examine the statistical assumptions underlying the analysis of variance for different response variables as measures of soft rot resistance using the tuber slice method of evaluation; 2) estimate the sample sizes necessary to detect specified differences with power 0.83; and, 3) choose the “best” variable for measuring resistance to soft rot based on valid statistical analysis and minimal sample size. Slices from fifteen tubers from each of three cultivars (Atlantic, Norchip, Superior) were inoculated withErwinia carotovora subsp.atroseptica, E. carotovora subsp.carotovora orE. chrysanthemi and incubated at two different temperatures (20 C and 30 C) for 48 hrs. The test was conducted on two dates. Tuber samples were sliced and weighed prior to inoculation and after the macerated tissue was removed following a 48 hr incubation period. The maximum diameter of macerated tissue, actual weight loss, four measures of proportional weight loss, and various transformations of these variables were analyzed. The “best” response variables for measuring resistance to soft rot by the tuber slice method were the diameter of the macerated tissue and the square root transformation of one of the measures of proportional weight loss. No differences were found among the cultivars nor theErwinia subsp. for either of these response variables, and the cultivar ×Erwinia subsp., cultivar × temperature and cultivar ×Erwinia subsp. × temperature interactions were not significant. However, there were significant differences between the incubation temperatures, and theErwinia subsp. × temperature interaction was significant for both response variables. Estimates of sample sizes necessary to find a 20% difference in main effect and interaction effect means were calculated.     

The Effect of Long Term Storage on Bacterial Soft Rot Resistance in Potato

Bacterial soft rot is a serious disease in potato (Solanum tuberosum L.), causing rapid tuber tissue maceration and, consequently, marketable yield loss. Soft rot bacteria, including Pectobacterium carotovorum subsp. carotovorum (Pbc), are favored by moist conditions, which are prevalent in large potato storage facilities. However, although most potatoes in North America are stored before use, there are no published surveys of soft rot resistance in cultivars exposed to long-term storage conditions. Thus, we tested 65 cultivars and 13 breeding lines for soft rot resistance after 6 months of storage. There was a significant effect of cultivar and production environment on soft rot resistance score. During 6 months of storage, tuber soft rot resistance in resistant clones did not change, while it changed in susceptible clones. The three most resistant cultivars to soft rot were Freedom Russet, Anett, and Alaska Red Eye.     

Suppression of bacterial soft rot in potato tubers by application of kasugamycin

An immersion in aqueous solutions of Kasumin®, containing kasugamycin at 10 to 320 mg/L (ppm) for 1 to 320 sec delayed or prevented the development of soft rot on inoculated (Erwinia carotovora subsp.carotovora) tuber disks incubated over wet filter paper at 24C for 24 to 72 hr. Treatment efficacy decreased as inoculum level and incubation period increased. Short immersion periods were not as effective as longer ones. When stored tubers were inoculated withE. c. carotovora and incubated up to 5 days in fog chamber at 20 C, immersion treatments in 20 to 400 ppm kasugamycin either had no effect or increased soft rot development. By contrast, when the stored tubers were cut into sections before inoculation and treatment, immersion in 300 ppm reduced soft rot development on the cut surfaces from 83% (inoculated controls) to 3.2%. With non-inoculated sections, treatment with 25 ppm reduced the SAD from 13.3 to 0.6%. Wash or rinse treatment of freshly harvested tubers in 40 to 160 ppm produced a significant reduction in the severity of bacterial soft rot among tubers incubated for 72 to 96-hr. The 160-ppm treatment delayed disease onset by at least 24 hr. Pre-washing freshly harvested tubers with clean or chlorinated water prior to antibiotic treatment did not consistently improve the efficacy of the kasugamycin treatment.     

Resistance of tubers from different potato cultivars to soft rot caused byErwinia carotovora subsp.atroseptica

Mechanically harvested tubers of 14 potato cultivars grown on both loamy sand and silt loam soils were evaluated for resistance to bacterial soft rot caused byErwinia carotovora subsp.atroseptica. Cultivars were also assayed for calcium and dry matter content to determine possible correlations with soft rot resistance. Resistance of potato tubers to bacterial soft rot was assayed after harvest by inoculating tubers and determining percent surface area decayed after four days in a mist chamber. Tubers of the cultivars differed widely in resistance to bacterial soft rot. The ranking of the resistance of tubers from different cultivars to bacterial soft rot was similar for both soils, although disease severity in tubers from a particular soil type varied with the season. Of the cultivars tested, tubers of Butte, Russet Burbank, and Nor-gold Russet were ranked as the most resistant, whereas tubers of Redsen, Norchip, Norland and Belchip were rated as the most susceptible. Tubers of Rhinered, Red La Soda, Superior, Pontiac, Atlantic, and Nooksack were intermediate in resistance. Neither calcium concentration in medullar tissues nor dry matter of potato tubers alone could be correlated with ranking of the cultivars with respect to severity of bacterial soft rot. However, a multiple linear regression model incorporating both variables indicated that tubers with a high content of both calcium and dry matter were likely to be less susceptible to bacterial soft rot than were tubers with lower percentages of these variables.     

Introgression and stabilization oferwinia tuber soft rot resistance into potato after somatic hybridization ofsolanum tuberosum ands. brevidens

Resistance to potato tuber soft rot caused byErwinia carotovora was transferred fromSolanum brevidens to the cultivated potato over the course of four backcross generations originating from a somatic hybrid. Soft rot reactions were determined via a tuber plug inoculation method developed during the course of these experiments. Soft rot resistance was highest in the somatic hybrid (only ca. 20% of tubers and plugs showed evidence of severe rotting) and lowest among progeny of control potato x potato crosses (ca. 80% of tuber plugs showed severe rot). Backcross generations involving somatic hybrids were intermediate in their reaction, and resistance stabilized to about 60% of tuber plugs showing severe rot in the BC₂ through the BC₄. Reciprocal crosses showed no difference in the inheritance of soft rot resistance, indicating that neitherS. brevidens norS. tuberosum donor cytoplasm had a significant effect on the expression of resistance. Crosses between BC₃ siblings where noS. brevidens genetic markers were detected but resistance was segregating demonstrated a dosage effect for soft rot resistance. We conclude that introgression of soft rot resistance has occurred and that at least one locus responsible for resistance inS. brevidens now resides in theS. tuberosum genome.     

Evaluation for chipping and tuber soft rot (Erwinia carotovora) resistance in potato clones from unilateral sexual polyploidization (2x × 4x)

The aim of this work is the evaluation of tetraploid potato clones with germplasm fromS. chacoense andS. phureja for chipping and resistance to tuber soft rot (Erwinia carotovora subsp.carotovora). Tetraploid potato clones were obtained via 2n eggs through 2x–4x crosses. Cytological mechanisms were investigated in the diploid hybrid 2n-egg producer used as parent in the unilateral sexual polyploidization scheme; the omission of the second meiotic division and cytokinesis failure at the end of meiosis were identified with a genetic consequence equivalent to second division restitution (SDR). Tuber marketable yield, specific gravity and chip-processing ability evaluated at harvest and after cold storage with and without reconditioning were higher in some clones as compared to theS. tuberosum tetraploid parent. Tuber soft rot resistance, contributed byS. chacoense, was successfully transferred to the4x level. After inoculation withErwinia carotovora subsp.carotovora and estimation of the rotted area, one tetraploid clone proved to be resistant and three partially resistant. The genetic value of exotic germplasm and sexual polyploidization strategy are discussed in relation to potato improvement.     

Soft-rot Resistance of Coloured Potato Cultivars (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.): The Role of Anthocyanins

Coloured potatoes (Solanum tuberosum L.) rich in anthocyanins are gaining increasing interest on the markets, mainly due to beneficial health effects. Potato varieties containing colour pigments are therefore utilized more and more in actual breeding programmes. In this study, 10 purple-fleshed potato cultivars were compared in their resistance to soft rot caused by Pectobacterium carotovorum (Pc) with 10 white/yellow-fleshed potato cultivars. The results revealed that coloured potato cultivars exhibited on average better resistance to soft rot than white/yellow-fleshed potato cultivars. On tuber halves of purple-fleshed cultivars, the extent of rotting caused by Pc was on average 28.6% lower than on white/yellow-fleshed cultivars. The greater resistance of coloured potatoes coincided with the presence of anthocyanins, higher concentrations of soluble phenols and elevated polyphenol oxidase activity in tuber tissue. These three components were significantly correlated with the extent of rotting caused by Pc. However, coloured and white/yellow-fleshed cultivars were not very different in phenylalanine ammonia-lyase and peroxidase activity. There were also no significant differences in concentrations of dry matter, starch, crude protein and glycoalkaloids. Key findings reveal that total soluble phenols and anthocyanins contributed crucially to resistance expression in tuber tissue of coloured potato cultivars.     

Quaggy Joe: A high-yielding, round, white potato variety for fresh market

The Quaggy Joe potato variety is high-yielding and medium to medium-late maturing with attractive, round-oval, white-skinned, white-fleshed tubers with medium-shallow pink eyes. Its major use is expected to be as a maincrop tablestock variety. Quaggy Joe is resistant to the potato leafroll virus, and is also moderately resistant to early blight (Alternaria solani), dry rot (Fusarium sambucinum), and soft rot (Erwinia carotovora subsp.carotovora). Quaggy Joe tubers have shown very little shatter or blackspot bruising, but sprout earlier than Atlantic. Young plants have shown sensitivity to the herbicide metribuzin.     

Screening tuber-bearingSolanum spp. for resistance to soft rot caused byErwinia carotovora ssp.atroseptica (van Hall) Dye

Summary Soft rot caused by Erwinia carotovora ssp. atroseptica is a major disease of stored potatoes. Since varietal resistance can contribute to control, the work reported was designed to find new sources of resistance among related tuber-bearing Solanum spp. True seeds were imported from two international collections and families were screened for resistance to tuber soft rot. Forty-eight resistant clones were found in 21 out of 100 accessions. These clones will be used in breeding programmes at the diploid or tetraploid level.     

Assessing six Austrian potato cultivars for resistance toErwinia carotovora subsp.atroseptica

Summary The susceptibility of six new Austrian cultivars to soft rot, stem rot and blackleg caused byErwinia carotovora subsp.atroseptica was assessed over 2 years in two locations. Soft rot susceptibility was assessed by whole tuber and half tuber tests, stem rot by stem inoculation and blackleg by planting inoculated tubers in the glasshouse. The susceptibility of the cultivars tested with the tuber and blackleg test methods were in general agreement. The half tuber test was more precise in differentiating intermediate levels of susceptibility than the whole tuber test. The reaction of the cultivars to soft rot could be classified as susceptible (Gina and Romina), moderately susceptible (Ditta, Sonja and Bintje), or slightly susceptible (Treff, Komet and Désirée). Results from the stem rot test did not agree with this classification and also depended upon the physiological state of plants as influenced by growing season and glasshouse conditions.     

Identification of stem soft rot (Erwinia carotovora subspeciesatroseptica) resistance in potato

Development of potato cultivars resistant to Erwinia species (Erwinia carotovora subspeciesatroseptica, E. carotovora subspeciescarotovora, andErwinia chrysanthemi) causing soft rot of stems and tubers involves identification and transfer of the resistance to breeding lines possessing desirable agronomic traits. In total, 363 accessions of six wildSolanum species, three somatic hybrids and nine sexual progeny of somatic hybrids were screened for stem soft rot resistance. Thirty-one, 50, and 67 percent of accessions ofSolanum boliviense, Solanum chacoense, and Solanumsanctarosae, respectively, were resistant or highly resistant, whereas, 100, 97, and 95 percent of accessions ofSolanum canasense, Solanum tarijense, andSolanum spegazzinii were susceptible or highly susceptible, respectively. Two tuber soft-rot resistant somatic hybrids, A937 and T355-11 produced earlier by the fusion ofSolanum brevidens (PI 218228) andSolanum tuberosum (PI203900) orS. tuberosum cv. Russet Burbank, respectively, and the sexual progeny of A937 andS. tuberosum cv. Katahdin were also highly resistant. Transfer of resistance fromS. brevidens to somatic hybrid (A937) and subsequently to the progeny suggests that the resistance is simply inherited. In addition to the identification of 65 resistant or highly resistant clones of wildSolanum species, this study has identified three tuber soft rot-resistant clones that are also resistant to stem soft rot. Potato cultivars showed a range of responses to the pathogen.     

Susceptibility of potatoes to bacterial rot and weight loss as a function of wound-healing interval and temperature

A study was conducted to determine the feasibility and advantages of wound-healing potatoes at lower than recommended temperatures. The progress of wound-healing at 6, 9, and 16°C, of tissue taken from Russet Burbank potato tubers, was evaluated by inducing weight loss with high temperature or decay organisms following various curing intervals. Increasing the wound-healing temperature resulted in a greater resistance to weight loss and a decreased susceptibility to infection byErwinia carotovora subsp.carotovora. Resistance to weight loss increased as wound-healing temperature increased following 6 and 16 day intervals. However, weight loss induced after 21 days of healing appeared to level off at 9°C indicating maximum development of a wound barrier. Loss due to rot, after inoculation of fresh cores withE. carotovora, was directly proportional to incubation time and temperature. When wound-healing progress was measured by loss of fresh weight induced by elevated temperatures or rot, the advantages of woundhealing at 16°C compared with 9°C decreased significantly after 21 days of curing. Wound-healing at approximately 9°C for 25 days was found to be the best compromise temperature for keeping rot progress and weight loss at a minimum while allowing the process of healing and maturation to occur.     

Effect of acetylsalicylic acid on soft rot produced byErwinia carotovora subsp.carotovora in potato tubers under greenhouse conditions

Summary The efficiency of acetylsalicylic acid (ASA) in inducing localised acquired resistance against infection byErwinia carotovora subsp.carotovora was evaluated by treating potato tubers with ASA at three concentrations. Three days after treatments, tubers were inoculated withE. carotovora subsp.carotovora by wound inoculation or by irrigation with a bacterial suspension. Experiments were performed for two consecutive years. Statistical analysis revealed that treatment of tubers by immersion in ASA solutions at low concentrations induced a significant reduction in the soft rot incidence. Wounding of the tubers was the most effective inoculation method and ASA at concentration of 0.0125% (w/v), pH 7 was more efficient than at 0.025 or 0.05%. No phytotoxicity of such treatment was observed.     

Calcium and potassium fertilization of potatoes grown in North Florida. II. Effect on the bacterial soft rot potential in the tubers

In studies on a sandy soil in Florida over a 3-yr period (1984–86), applications of Ca (CaSO₄) inconsistently affected the bacterial soft rot potential of potato tubers (Solanum tuberosum). The potential was estimated from the severity of the disease (percent surface area decayed) in tubers that had been immersed in an aqueous cell suspension ofErwinia carotovora subsp.carotovora and then incubated 4 days in a mist chamber at 24 C. With freshly harvested tubers (tested within 5 days after harvest), disease severity averaged 49, 11, and 4% over all treatments in 1984, 1985, and 1986, respectively, which was roughly proportional to rainfall on the crop,e.g., 37, 18, and 15 cm, respectively. Disease severity decreased, did not change, and increased with increased Ca rate (0, 450, and 900 kg.ha^-1) in 1984, 1985, and 1986, respectively. In each season, severity was affected by interactions involving the Ca treatments. Most interactions involved cultivar. The efficacy of Ca applications to potatoes grown in low Ca soils for enhancement of tuber resistance to bacterial soft rot may be limited by factors associated with environment or cultivar that are not completely understood.     

Resistance to blackleg and tuber soft rot in sexual and somatic interspecific hybrids with different genetic background

Diploid interspecific hybrids from crosses betweenSolanum tuberosum haploids andS. canasense, S. multidissectum, andS. tarijense, along with tetraploid and hexaploid somatic hybrids S.tuberosum (+)S. commersonii, were screened for resistance to blackleg and tuber soft rot byErwinia carotovora subsp.atroseptica andE. carotovora subsp.carotovora. Among the diploid hybrids, those involving clone mlt la of S.multidissectum and tar 2b ofS. tarijense yielded the highest number of genotypes resistant to both blackleg and tuber soft rot. By contrast, all the hybrids involving clone tar 1lb ofS. tarijense were susceptible to both bacterial diseases. As far as resistance of somatic hybrids is concerned, the most interesting genotype was the tetraploid one which showed resistance to both blackleg and tuber soft rot Significant correlations were found betweenE. carotovora subsp.atroseptica andE. carotovora subsp.carotovora both in resistance to blackleg and in resistance to tuber soft rot, suggesting that resistance to the two subspecies is controlled by the same gene/s or that the relevant genes are linked. On the other hand, correlations between blackleg and tuber soft rot were never significant, indicating that different mechanisms may control resistance in tuber and stems. Alternatively, the resistance could be controlled by the same gene/s under a different spatial/temporal expression pattern. Twelve diploid sexual hybrids and one tetraploid somatic hybrid were selected for resistance to blackleg and/or tuber soft rot and for tuber characteristics.     

Enhanced resistance to bacterial infection byErwinia carotovora subsp.atroseptica in transgenic potato plants expressing the attacin or the cecropin SB-37 genes

Blackleg and soft rot diseases, caused by the bacteriumErwinia carotovora, are among the diseases that cause important losses in culture and storage of potato. In this paper, we introduced bacterial resistance into potato, via genes encoding for proteins with antibacterial activity. For this purpose, potato clones were transformed either with the gene encoding the acidic attacin protein fromHyalophora cecropia, or with the gene encoding the cecropin analog peptide SB37. These clones were evaluated for soft rot and blackleg resistance, after inoculation with the bacterial strainErwinia carotovora subsp.atroseptica T7. Results reported in this paper indicate that a considerable percentage of the potato clones (15–22%) showed increased resistance to bacterial infection, revealed by reduced severity of blackleg or soft rot symptoms. Expression of the transgenes was demonstrated in some of the clones by Northern blot analysis. This is the first report indicating that expression of the gene encoding for an attacin protein and for the cecropin SB-37 peptide in transgenic potato confers increased resistance to bacterial infection.