Evaluation of potato tuber moth (Lepidoptera: Gelechiidae) resistance in tubers of Bt-cry5 transgenic potato lines

The potato tuber moth, Phthorimaea operculella (Zeller), in tropical and subtropical countries, is the most destructive pest of potato, Solanum tuberosum L. The larvae attack foliage and tubers in the field and in storage. The purpose of this study was to evaluate the efficacy of a Bt-cry5 transgene to control the potato tuber moth in tuber tissues. Tuber bioassays using stored (11-12 mo old) and newly harvested tubers of Bt-cry5-Lemhi Russet and Bt-cry5-Atlantic potato lines showed up to 100% mortality of 1st instars. Mortality was lowest in the newly harvested tubers of Bt-cry5-Atlantic lines (47.1-67.6%). Potato tuber moth mortality was 100% in the Bt-cry5-Spunta lines that were transformed with Bt-cry5 gene controlled by the CaMV 35S promoter (pBIML5 vector) and in 2 of 3 lines transformed with Bt-cry5 gene controlled by the Gelvin super promoter (pBIML1 vector). The transgenic Spunta lines expressing Bt-cry5 controlled by the patatin promoter (pBMIL2 vector) showed the lowest tuber moth mortality (25.6 and 31.1%). The Bt-cry5 transgenic lines with high tuber expression of B. thuringiensis have value in an integrated pest management system to control potato tuber moth.     

Effects of gamma irradiation used to inhibit potato sprouting on potato tuber moth eggs Phthorimaea operculella Zeller (Lep., Gelechiidae)

Abstract: Different age groups of potato tuber moth, Phthorimaea operculella Zeller, eggs were exposed to gamma irradiation at incremental doses up to 150 Gy (the upper dose limit allowed for potato sprout inhibition). Young eggs were more likely to be sensitive to gamma irradiation than older eggs and the sensitivity level declined with age of the eggs. The exposure of 3–3.5-day-old eggs to 150 Gy resulted in a 26% increase in the egg incubation period compared with unirradiated eggs. The minimum dose required to prevent 4–4.5-day-old eggs from hatching was 1 kGy. When eggs were exposed to 75 Gy, 9.7% of larvae survived to the adult stage but emerged as deformed moths. At 100 and 125 Gy, 10.2 and 9.6% of larvae pupated, respectively, although no adult eclosion was recorded, whereas at 150 Gy, the larvae remained in mid-instar stage and eventually died. Gamma irradiation doses applied to inhibit potato sprouting could be also considered as a valuable control tool against potato tuber moth infestations.     

Biological Control of the Potato Tuber Moth (Phthorimaea operculella Zeller) in the Republic of Yemen Using Granulosis Virus: Biochemical Characterization, Pathogenicity and Stability of the Virus

A granulosis virus infecting the potato tuber moth, Phthorimaea operculella, has been identified, isolated and purified from diseased potato tuber moth larvae collected from potato fields in the highland basin Qa al-Boun of the Republic of Yemen. The granulosis virus was propagated by feeding potato tuber moth larvae with potatoes treated with a concentration of one granulosis virus-infected fourth instar larva to 5l of water (corresponding to an occlusion body (OB) concentration of 2 106 OB ml-1). Restriction enzyme analysis of viral DNA revealed that the isolated virus seemed to correspond to an isolate from the Lima region of Peru. A median LC 50 value of 7.3 104 OB ml-1 was calculated for a purified virus preparation. Different preparations of the granulosis virus were investigated for their persistence in the field on tubers and leaves. A purified virus preparation (PoGV) applied to potato tubers and exposed in the open had a half-life of 1.3 days. On leaves, the activity of granulosis virus spray deposits of an unpurified virus preparation (PoGV I) and of a glycerine-formulated preparation (PoGV II) also declined with exposure time. Mortalities of potato tuber moth larvae of 43% for PoGV I and 49% for PoGV II were recorded when first instar larvae were fed with potato leaves collected 2 days after treatment. After 8 days, 25.8% of the larvae had died from PoGV I treatment and 19.4% from PoGV II. Neither preparation displayed any effect 17 days after application.     

Spatial and temporal dynamics of potato tuberworm (Lepidoptera: Gelechiidae) infestation in field-stored potatoes

The potato tuberworm, Phthorimaea operculella (Zeller), is a major pest of potatoes in fields and traditional storage. A common method of nonrefrigerated storage is to pile potatoes in straw-covered heaps in the field. Tubers may be stored up to 3-4 mo in this manner, until the next harvest. We studied the dynamics of potato tuber moth infestation associated with such field storage in a 12-wk experiment in Israel. We set up six potato heaps, and sampled them for potato tuber moth at different locations at weekly intervals. Potato tuber moth infestation was significantly higher at the perimeter of the heaps than at their center, but it did not differ between bottom, mid-height, and top of the heaps. The proportion of potato tuber moth-infested potato tubers increased from 10 to 65% over the course of the experiment, and the mean number of potato tuber moth larvae per tuber increased from 0.25 to 2.50. Potato tuberworm populations increased sharply after 3, 6, and 9 wk of study, possibly corresponding to successive generations that developed within the heaps. This interpretation is supported by calculations of potato tuberworm generation length based on temperature data. Catches in pheromone traps that were placed near the heaps were not correlated (spatially and temporally) with potato tuberworm densities within heaps, hinting that migration among heaps did not considerably affect within-heap population dynamics. Potato tuberworm levels were not correlated with ambient temperatures, perhaps because of the warm, humid, and constant microclimate within the heaps. We discuss the significance of our findings for control efforts of the potato tuberworm.     

Resistance of potatoes transgenic for a cry1Ac9 gene, to Phthorimaea operculella (Lepidoptera: Gelechiidae) over field seasons and between plant organs

Stable performance of insect‐resistant transgenic plants across field seasons and between plant organs damaged by the insect pest is critical for management of this resistance in the field. To evaluate this, potato (Solanum tuberosum) lines transgenic for a cry1Ac9 gene with resistance to potato tuber moth (Phthorimaea operculella) were established in the field during the southern hemisphere summers of 1997/98, 1998/99 and 1999/00 as small field plots, each of 10 plants. Replicate plots of the non‐transgenic parent cultivars (at least one for every three independently derived transgenic lines) were planted randomly throughout the trials. Field‐grown foliage was challenged with larvae in the laboratory and a growth index (GI) was calculated for recovered larvae from each transgenic and non‐transgenic potato line. Larval growth on young and mature leaves, and on newly harvested or stored tubers was also measured in the laboratory. Foliage from the transgenic lines inhibited larval growth in all seasons tested. For both control and transgenic lines, larvae had slightly lower GIs when reared on mature leaves compared with young leaves, although the correlation between mean GI for young and mature transgenic leaves was high (r = 0.97). The correlation between the mean GIs of larvae on newly harvested tubers and on those stored for 5 months was also high (r = 1.0). However, the GIs of larvae on newly harvested transgenic tubers were larger than on transgenic tubers stored for 5 months. The relative growth indices (RGI = mean GI/number days before final weighing) of larvae reared on newly harvested tubers from transgenic lines were generally higher than those from young transgenic foliage, while the RGIs of larvae reared on non‐transgenic tubers were slightly lower than those fed non‐transgenic foliage. The correlation between mean RGIs of larvae fed tubers or foliage was 0.62. The transgenic potato lines exhibited stable resistance to larvae across field seasons, between affected plant organs, and between plant organs of different ages.     

Susceptibility of potato tuber moth (Lepidoptera: Gelechiidae) to postharvest gamma irradiation

Gamma irradiation doses applied to inhibit potato sprouting were tested against potato tuber moth larval and pupal stages. Young larvae and pupae were more susceptible to irradiation injuries than older ones. When larvae of different ages were exposed to doses > or = 100 Gy, only 13-35% pupated, but no adult emergence was obtained. The exposure of 1-1.5- or 3-3.5-d-old pupae to 150 Gy induced high level of sterility and remarkable reduction in female mating ability and fecundity, whereas the reduction was less noticeable for 5-5.5-d-old pupae. Postharvest gamma irradiation of potatoes could be considered as an efficient control approach against potato tuber moth larval and pupal infestations.     

用马铃薯人工饲养马铃薯块茎蛾的方法

介绍一种马铃薯块茎蛾Phthorimaeaoperculella的室内人工饲养方法。在人工养虫室内用 1 0 %的蜂蜜水喂养成虫 ,用滤纸收集卵块 ,幼虫在薯块上饲养 ,化蛹于牛皮纸卷成的纸筒中 ,饲养结果发现马铃薯块茎蛾完成 1代大约需要 35～ 4 6d ,室内连续饲养块茎蛾 3代 ,卵的孵化率、幼虫化蛹率、成虫羽化率均在 90 %以上。     

Age-specific bioassays and fecundity of Phthorimaea operculella (Lepidoptera: Gelechiidae) reared on cry1Ac -transgenic potato plants

Variation in the susceptibility of lepidopterous pest larvae of different ages to transgenic crops and the potential for survivors to reproduce could have important consequences for the development of resistance in such pests. Experiments were undertaken in the laboratory to determine if larvae of the potato tuber moth, Phthorimaea operculella, of different ages (0 (< 1 day old), 3, 5, 7 days) varied in their susceptibility to cry1Ac9–transgenic potato (Solanum tuberosum) foliage grown in the glasshouse or field. The survival and fecundity of larvae reared on transgenic tubers was also determined in the laboratory. There were no apparent differences in susceptibility of larvae of different ages to transgenic foliage. Larvae fed glasshouse or field‐grown non‐transgenic foliage had significantly larger relative growth indices and more larvae pupated, than those fed transgenic foliage, regardless of larval age. Eggs from a laboratory colony were placed on transgenic or non‐transgenic tubers to measure survival and fecundity. Between 6% and 15% of eggs placed on transgenic tubers developed into pupae for three of the four transgenic potato lines tested. On one transgenic line, only six adults emerged from 1300 eggs. In contrast, between 71% and 97% of the eggs placed on non‐transgenic tubers developed into pupae. Male and female pupae from transgenic lines weighed less than those from non‐transgenic lines. The fecundity of females from two of four transgenic lines was lower than from the non‐transgenic parent cultivar. Although larvae of different ages did not exhibit any overall age‐dependent pattern of increasing or decreasing susceptibility to transgenic foliage of glasshouse or field‐grown plants, the ability of larvae to survive and reproduce on transgenic tubers suggests this pest has the ability to evolve resistance to the transgenic plants used in the present study.     

Tecia solanivora infestation increases tuber starch accumulation in Pastusa Suprema potatoes

In response to infestation with larvae of the Guatemalan tuber moth(Tecia solanivora), some Solanum tuberosum(potato) varieties exhibit an overcompensation response, whereby the total dry mass of uninfested tubers is increased. Here, we describe early responses,within the first few days, of T. solanivora feeding, in the Colombian potato variety Pastusa Suprema. Nontargeted metabolite profiling showed significant secondary metabolism changes in T. solanivora-infested tubers,but not in uninfested systemic tubers. In contrast,changes in primary metabolism were greater in uninfested systemic tubers than in the infested tubers, with a notable 80% decline in systemic tuber sucrose levels within 1 d of T. solanivora infestation. This suggested either decreased sucrose transport from the leaves orincreased sink strength, i.e., more rapid sucrose to starch conversion in the tubers. Increased sucrose synthesis was indicated by higher rubisco activase and lower starch synthase gene expression in the leaves of infested plants.Elevated sink strength was demonstrated by 45% more total starch deposition in systemic tubers of T. solanivorainfested plants compared to uninfested control plants.Thus, rather than investing in increased defense of uninfested tubers, Pastusa Suprema promotes deposition of photoassimilates in the form of starch as a response to T. solanivora infestation.     

Oviposition deterrence of shoots and essential oils of Minthostachys spp. (Lamiaceae) against the potato tuber moth

Abstract:  The potato tuber moth is a noxious pest of potato in stores, where the use of repellent plants is an environmentally sound alternative to the application of chemical pesticides. We evaluated the protective effect of native Minthostachys species (Lamiaceae) against tuber infestation by the potato tuber moth in a rustic store in Cusco, Perú. We covered potato tubers with dried shoots of Minthostachys spicata and Minthostachys glabrescens and compared tuber damage with a control treatment of maize straw. We also conducted a no-choice oviposition bioassay in the laboratory, testing the oviposition deterrence of essential oils of M. spicata , M. glabrescens and Minthostachys mollis at natural concentrations. We recorded the number of eggs laid by mated moths on filter paper treated with essential oils of each of the three species and on two control treatments: hexane and untreated blank. Finally, we tested for differences in oviposition deterrence among five full-sib families of potato tuber moth raised under identical conditions. We found that dried, chopped leaves and flowers of Minthostachys species reduced the percentage of tuber damage in stores in comparison with the control (5% vs. 12%), but no difference in protection was found between species. Essential oils at natural concentrations deterred moth oviposition, reducing the number of eggs laid by about 80% compared with the control treatments; again, there were no significant differences between Minthostachys species. Finally, whereas we detected among-family variation in oviposition on filter papers treated with essential oils, no difference was found in the number of eggs laid on control substrates. Therefore, there was genetic variation for oviposition deterrence in the potato tuber moth and resistance to repellent plants might evolve thereafter.     

Laboratory evaluation of lufenuron on immature stages of potato tuber moth (Lepidoptera: Gelechiidae)

The effects of the chitin synthesis inhibitor lufenuron against potato tuber moth, Phthorimaea operculella (Zeller), eggs were determined by topically exposing different age groups of eggs (1-4 d old) to treated potato tubers (Solanum tuberosum L.) under laboratory conditions. Larval hatch from both treated (4 and 12 g [AI]/100 liter) and untreated tubers was >95%, but mortality of first instars was high in treated tubers (>90%) compared with untreated tubers. Examination of the treated tubers showed that most of the larvae were unable to penetrate or cause any noticeable damage to the potato tubers. However, the few first instars that survived were able to penetrate the tubers and continue their development to the pupal or adult stages. At 12 g (AI)/100 liter, adult emergence was <2% and most of the emerged adults had morphological deformities such as reduced wing size and they were unable to free themselves from the pupal sacs. These data suggest that topical application of lufenuron to eggs before larval hatch would reduce the amount of damage caused by potato tuber moth as part of integrated pest management program.     

Variations in resistance against Phthorimaea operculella in wild potato tubers

Tuber resistance can contribute to current management strategies against the potato tuber moth, Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae), in field and stored potatoes. Wild potatoes represent a potential source of novel resistance traits against the moth. We assessed resistance in three wild potato species, Solanum multiinterruptum Bitt., Solanum sparsipilum (Bitt.) Juz. & Buk., and Solanum wittmackii Bitt. against neonate and developing tuber moth larvae. All three species had high levels of resistance but accessions of S. sparsipilum and S. wittmackii were significantly more resistant. Resistance in S. multiinterruptum was generally concentrated in the tuber periderm, whereas in S. sparsipilum and S. wittmackii resistance was mainly cortex‐based. Unidentified cortex‐resistance factors in all three species reduced survival and increased larval and pupal development times, but had no apparent effects on the pupal weights of survivors. A high proportion of larvae abandoned or died within tubers of S. wittmackii, which has particularly high levels of unidentified cortex‐based defenses. Resistance decreased in S. multiinterruptum and S. sparsipilum as tubers sprouted but was more stable in S. wittmackii. Periderm‐based resistance was more stable than cortex‐based resistance in S. multiinterruptum during sprouting. In contrast, cortex‐based resistance was stable in tubers of S. wittmackii as these sprouted, and resistance may have increased on some older sprouting tubers. Solanum multiinterruptum and S. sparsipilum are proposed as potential sources of resistance against the potato tuber moth.     

Repression of a novel isoform of disproportionating enzyme (stDPE2) in potato leads to inhibition of starch degradation in leaves but not tubers stored at low temperature

A potato (Solanum tuberosum) cDNA encoding an isoform of disproportionating enzyme (stDPE2) was identified in a functional screen in Escherichia coli. The stDPE2 protein was demonstrated to be present in chloroplasts and to accumulate at times of active starch degradation in potato leaves and tubers. Transgenic potato plants were made in which its presence was almost completely eliminated. It could be demonstrated that starch degradation was repressed in leaves of the transgenic plants but that cold-induced sweetening was not affected in tubers stored at 4 degrees C. No evidence could be found for an effect of repression of stDPE2 on starch synthesis. The malto-oligosaccharide content of leaves from the transgenic plants was assessed. It was found that the amounts of malto-oligosaccharides increased in all plants during the dark period and that the transgenic lines accumulated up to 10-fold more than the control. Separation of these malto-oligosaccharides by high-performance anion-exchange chromatography with pulsed-amperometric detection showed that the only one that accumulated in the transgenic plants in comparison with the control was maltose. stDPE2 was purified to apparent homogeneity from potato tuber extracts and could be demonstrated to transfer glucose from maltose to oyster glycogen.     

Zebra chip disease incidence on potato is influenced by timing of potato psyllid infestation, but not by the host plants on which they were reared

Abstract  The Zebra chip (ZC) syndrome is an emerging disease of potato and a major threat to the potato industry. The potato psyllid, Bactericerca cockerelli (Sulc) is believed to be a vector of the ZC pathogen, which is now thought to be Candidatus Liberibacter, a bacterium. To further understand the relationship between potato psyllid infestation and ZC disease expression, healthy potato plants at different growth stages (4, 6 and 10 weeks after germination) were exposed separately to potato psyllids that were separately reared on four solanaceous hosts plants (potato, tomato, eggplant or bell pepper) for more than 1 year. ZC symptoms, leaf rates and total nonstructural carbohydrate accumulation in leaves and tubers of healthy and psyllid-infested plants were monitored and recorded. Typical ZC symptoms were observed in leaves and tubers of all plants exposed to potato psyllids regardless of the host plant on which they were reared. This was also accompanied by significant reductions in net photosynthetic rate. Caged potato plants without exposure to potato psyllids (uninfested controls) did not show any ZC symptom in both foliage and in harvested tubers. Foliage damage and ZC expression were most severe in the potato plants that were exposed to potato psyllids 4 weeks after germination compared to plants infested at later growth stages. Tubers from potato psyllid-infested plants had significantly higher levels of reducing sugars (glucose) and lower levels of starch than those in healthy plants, indicating that potato psyllid infestation interfered with carbohydrate metabolism in either leaves or tubers, resulting in ZC expression.     

The three-dimensional distribution of minerals in potato tubers

Background and Aims

The three-dimensional distributions of mineral elements in potato tubers provide insight into their mechanisms of transport and deposition. Many of these minerals are essential to a healthy human diet, and characterizing their distribution within the potato tuber will guide the effective utilization of this staple foodstuff.

Methods

The variation in mineral composition within the tuber was determined in three dimensions, after determining the orientation of the harvested tuber in the soil. The freeze-dried tuber samples were analysed for minerals using inductively coupled plasma-mass spectrometry (ICP-MS). Minerals measured included those of nutritional significance to the plant and to human consumers, such as iron, zinc, copper, calcium, magnesium, manganese, phosphorus, potassium and sulphur.

Key Results

The concentrations of most minerals were higher in the skin than in the flesh of tubers. The potato skin contained about 17 % of total tuber zinc, 34 % of calcium and 55 % of iron. On a fresh weight basis, most minerals were higher in tuber flesh at the stem end than the bud end of the tuber. Potassium, however, displayed a gradient in the opposite direction. The concentrations of phosphorus, copper and calcium decreased from the periphery towards the centre of the tuber.

Conclusions

The distribution of minerals varies greatly within the potato tuber. Low concentrations of some minerals relative to those in leaves may be due to their low mobility in phloem, whereas high concentrations in the skin may reflect direct uptake from the soil across the periderm. In tuber flesh, different minerals show distinct patterns of distribution in the tuber, several being consistent with phloem unloading in the tuber and limited onward movement. These findings have implications both for understanding directed transport of minerals in plants to stem-derived storage organs and for the dietary implications of different food preparation methods for potato tubers.     

连作马铃薯植株库源关系及其对块茎产量的调节机理

甘肃中部沿黄灌区是国内重要的加工型马铃薯生产基地和种薯繁殖基地,但集约化生产带来的连作障碍已严重影响到产业的健康发展.为揭示马铃薯连作障碍机理,本研究通过田间试验,设置马铃薯连作年限为0～5年的处理,研究了马铃薯植株库容量、库活性和源活性对连作的响应特征及其对块茎产量的调节机制.结果表明: 短期连作(1～2年)条件下块茎产量较非连作(0年)无显著变化,长期连作(3～5年)显著下降28.6%～32.8%,单薯质量降低是导致块茎产量下降的直接原因.长期连作马铃薯库容量较非连作显著降低38.4%～53.0%.长期连作导致块茎形成推迟,同时单薯干物质积累量降低.长期连作也显著降低马铃薯源活性,与非连作相比,株高、主茎分枝数、叶绿素含量和叶片干物质量均显著下降,根系形态发育受到抑制,根系活力显著下降28.6%～63.1%,叶片RuBP羧化酶和蔗糖磷酸合成酶活性分别显著下降52.6%～64.6%和26.3%～53.4%.长期连作条件下马铃薯源端生产性能降低导致同化产物减少,花后阶段向块茎的输入不足,降低块茎产量.库源关系失衡是甘肃中部沿黄灌区长期连作马铃薯产量大幅降低的原因.     

Polyphenol oxidase in potato. A multigene family that exhibits differential expression patterns.

Polyphenol oxidase (PPO) activity in potato (Solanum tuberosum) plants was high in stolons, tubers, roots, and flowers but low in leaves and stems. PPO activity per tuber continued to increase throughout tuber development but was highest on a fresh weight basis in developing tubers. PPO activity was greatest at the tuber exterior, including the skin and cortex tissue 1 to 2 mm beneath the skin. Flowers had high PPO activity throughout development, particularly in the anthers and ovary. Five distinct cDNA clones encoding PPO were isolated from developing tuber RNA. POT32 was the major form expressed in tubers and was found in all parts of the tuber and at all stages of tuber development. It was also expressed in roots but not in photosynthetic tissues. POT33 was expressed in tubers but mainly in the tissue near the skin. POT72 was detected in roots and at low levels in developing tubers. NOR333 was identical with the P2 PPO clone previously isolated from potato leaves (M.D. Hunt, N.T. Eannetta, Y. Haifeng, S.M. Newman, J.C. Steffens [1993] Plant Mol Biol 21: 59-68) and was detected in young leaves and in tissue near the tuber skin but was highly expressed in flowers. The results indicate that PPO is present as a small multigene family in potato and that each gene has a specific temporal and spatial pattern of expression.     

Establishment of Beauveria bassiana as a fungal endophyte in potato plants and its virulence against potato tuber moth,Phthorimaea operculella (Lepidoptera: Gelechiidae)

The potato tuber moth, Phthorimaea operculella, is the most damaging potato pest in the world and is difficult to control as the larvae are internal feeders in the foliage and tubers. Entomopathogenic fungi that colonize plants as endophytes have lethal and sublethal pathological effects on insect pests. We show that Beauveria bassiana colonizes the aerial parts of potato plants endophytically after inoculation through soil drenching. Endophytic B. bassiana persisted in potato foliage for more than 50 days postinoculation. Bioassays indicated that foliage of B. bassiana-inoculated potato plants were pathogenic against larvae of P. operculella. Sublethal experiments indicated that B. bassiana negatively affected the growth, development, and reproduction of P. operculella. Development experiments showed that the weight of P. operculella pupae reared on B. bassiana-colonized potato plants (4.25 mg) was significantly less than that of those reared on uninoculated control plants (8.89 mg). Compared with newly eclosed larvae fed on control plants, those fed on B. bassiana-inoculated plants had significantly lower survivorship, with only 17.8% developing to the adult stage. Oviposition of P. operculella females reared on B. bassiana endophytically colonized plants was significantly lower (35 eggs/female) than of those reared on uninoculated plants (115 eggs/female). This study demonstrates that endophytic B. bassiana can be a potential biological control agent for the control and management of P. operculella. Comparing pupal weights of P. operculella reared on potato plants inoculated with the B. bassiana strain GZGY-1-3 and on untreated control plants, pupae from the control plants were significantly heavier than those from treated plants.