A transgenic study on affecting potato tuber yield by expressing the rice sucrose transporter genes OsSUT5Z and OsSUT2M

In many plants, sucrose transporters are essential for both sucrose exports from sources and imports into sinks, indicating a function in assimilate partitioning. To investigate whether sucrose transporters can improve the yield of starch plant, potato plants (Solanum tuberosum L. cv. Désirée) were transformed with cDNAs of the rice sucrose transporter genes OsSUT5Z and OsSUT2M under the control of a tuber-specific, class-I patatin promoter. Compared to the controls, the average fructose content of OsSUT5Z transgenic tubers significantly increased. However, the content of the sugars and starch in the OsSUT2M transgenic potato tubers showed no obvious difference. Correspondingly, the average tuber yield, average number of tubers per plant and average weight of single tuber showed no significant difference in OsSUT2M transgenic tubers with controls. In the OsSUT5Z transgenic lines, the average tuber yield per plant was 1.9-fold higher than the controls, and the average number of tubers per plant increased by more than 10 tubers on average, whereas the average weight of a single tuber did not increase significantly. These results suggested that the average number of tubers per plant showed more contribution than the average weight of a single tuber to the tuber yield per plant.     

Healthier seed potatoes. I. Effects of inoculating stem cutting stocks with Polyscytalum pustulans and Rhizoctonia solani on growth, yield and disease

Potato seed tubers of seven cultivars derived from stem cuttings in 1965 (healthier seed) were grown in 1969–72 at two sites, one clay with flints soil (Rothamsted) and the other sandy loam soil (Woburn). Inoculating sprouted tubers at planting with Polyscytalum pustulans did not affect the number of stems/plant or total yield but increased stem base and tuber infection. The yields of large tubers (57–83 cm) were increased and small tubers (>57 cm) decreased, indicating a decrease in tuber numbers. Rhizoctonia solani inoculated at planting decreased numbers of stems/plant and yield by up to 14% at Woburn but not at Rothamsted. At both sites, yields of large tubers were increased with cvs Majestic and Record and decreased with Pentland Crown. Stem canker and tuber infection were increased but infection was also prevalent on tubers from non-inoculated seed at Woburn. When both pathogens were inoculated together yields of large tubers were increased in cvs King Edward, Majestic and Record and decreased in Pentland Crown. Infection of stem bases and tubers was sometimes less than when either pathogen was inoculated singly.     

Effects of fluorescent pseudomonads on the potato blackleg syndrome

In four field trials in 1983 and 1984 potato tubers were inoculated by dipping them at planting in a suspension of Erwinia carotovora subsp. atroseptica and then treated with a powder formulation of two strains of fluorescent pseudomonads (B 10 and I 13) isolated from potatoes. lnoculation with E. carotovora increased blackleg and reduced emergence, plant growth, tuber size and weight compared with uninoculated controls. These effects were partially reversed by treatment of tubers with fluorescent pseudomonads which also reduced contamination by E. carotovora and the soft-rot potential of progeny tubers. In some trials a mixture of both pseudomonad isolates delayed the breakdown of the mother tuber although individual treatments did not.     

Growth and yield of potato plants inoculated with rhizosphere bacteria

Pot and field experiments on small plots were carried out to study the effect of isolates ofPseudomonas fluorescens-putida bacteria, applied to potato tubers, on the growth response of potato plants. Inoculation of tuber pieces with the isolates in pot experiments caused a better growth of young potato plant (111 % increase). The effect depended on the viable count in the bacterial suspension and on growth conditions of the plants. Inoculation of potato seed pieces before planting in field experiments caused a 4 –30 % improvement in plant growth and tuber yield. The complex action of inoculation with rhizobacteria apparently caused changes in the microflora colonizing the roots and stolons, which in turn brought about a better growth and yield.     

An analysis of the effects of nutrient supply on the growth of potato crops

Fertilizer nitrogen did not delay tuber initiation by the potato variety King Edward in the field, but slowed the early growth of tubers. Later, N hastened tuber growth and the largest amounts prolonged it. Leaf and stem growth were increased by N from an early stage. Leaf area index (L) reached maxima of 2.5 to 3.0 with the largest amounts of N, and 1.0 with no nitrogen: L decreased after late July, faster with increased N, and only the largest amount prolonged the life of the haulm. N increased leaf area duration (D) by up to 125% in both years. D was 21% greater in 1964 than 1963, and net assimilation rate in August and September was larger; consequently mean yield was 50% more in 1964. Uptake of nitrogen and N % of dry matter were increased by increasing nitrogen. The N % of tuber dry weight remained constant or increased slightly from about 2 weeks after tuber formation, while N % of the leaves and stems decreased rapidly, and much N was transferred from tops to tubers. The 1964 results suggest that tuber growth depends on continued nitrogen uptake by the plant. P and K had small effects compared with those of N. K increased leaf area duration by 9% in 1963 and 3% in 1964, and yield by 11 and 8% respectively. P increased leaf area duration by 17% and yield by 9% in 1964. K had very small effects until the end of the season, when it prolonged the period of growth. P increased growth rates of all parts of the plant up to 4 weeks from emergence, and the differences in dry weight at that time persisted to maturity but did not increase.     

Response of potato grown under non-inductive condition paclobutrazol: shoot growth, chlorophyll content, net photosynthesis, assimilate partitioning, tuber yield, quality, and dormancy

The effect of foliar and soil applied paclobutrazol on potato were examined under non-inductive condition in a greenhouse. Single stemmed plants of the cultivar BP1 were grown at 35(±2)/20(±2) °C day/night temperatures, relative humidity of 58%, and a 16 h photoperiod. Twenty-eight days after transplanting paclobutrazol was applied as a foliar spray or soil drench at rates of 0, 45.0, 67.5, and 90.0 mg active ingredient paclobutrazol per plant. Regardless of the method of application paclobutrazol increased chlorophyll a and b contents of the leaf tissue, delayed physiological maturity, and increased tuber fresh mass, dry matter content, specific gravity, dormancy period of the tubers. Paclobutrazol reduced the number of tubers per plant. A significant interaction between rates and methods of paclobutrazol application were observed with respect to plant height and tuber crude protein content. Foliar application gave a higher rate of net photosynthesis than the soil drench. Paclobutrazol significantly reduced total leaf area and increased assimilate partitioning to the tubers. The study clearly showed that paclobutrazol is effective to suppress excessive vegetative growth, favor assimilation to the tubers, increase tuber yield, improve tuber quality and extend tuber dormancy of potato grown in high temperatures and long photoperiods.     

Overriding the co-limiting import of carbon and energy into tuber amyloplasts increases the starch content and yield of transgenic potato plants

Transgenic potato (Solanum tuberosum) plants simultaneously over-expressing a pea (Pisum sativum) glucose-6-phosphate/phosphate translocator (GPT) and an Arabidopsis thaliana adenylate translocator (NTT1) in tubers were generated. Double transformants exhibited an enhanced tuber yield of up to 19%, concomitant with an additional increased starch content of up to 28%, compared with control plants. The total starch content produced in tubers per plant was calculated to be increased by up to 44% in double transformants relative to the wild-type. Single over-expression of either gene had no effect on tuber starch content or tuber yield, suggesting that starch formation within amyloplasts is co-limited by the import of energy and the supply of carbon skeletons. As total adenosine diphosphate-glucose pyrophosphorylase and starch synthase activities remained unchanged in double transformants relative to the wild-type, they cannot account for the increased starch content found in tubers of double transformants. Rather, an optimized supply of amyloplasts with adenosine triphosphate and glucose-6-phosphate seems to favour increased starch synthesis, resulting in plants with increased starch content and yield of tubers.     

Tuber-specific cphA expression to enhance cyanophycin production in potatoes

The production of biodegradable polymers that can be used to substitute petrochemical compounds in commercial products in transgenic plants is an important challenge for plant biotechnology. Nevertheless, it is often accompanied by reduced plant fitness. To decrease the phenotypic abnormalities of the sprout and to increase polymer production, we restricted cyanophycin accumulation to the potato tubers by using the cyanophycin synthetase gene ( cph A_Te) from Thermosynechococcus elongatus BP-1, which is under the control of the tuber-specific class 1 promoter (B33). Tuber-specific cytosolic (pB33- cph A_Te) as well as tuber-specific plastidic (pB33-PsbY- cph A_Te) expression resulted in significant polymer accumulation solely in the tubers. In plants transformed with pB33- cph A_Te, both cyanophycin synthetase and cyanophycin were detected in the cytoplasm leading to an increase up to 2.3% cyanophycin of dry weight and resulting in small and deformed tubers. In B33-PsbY- cph A_Te tubers, cyanophycin synthetase and cyanophycin were exclusively found in amyloplasts leading to a cyanophycin accumulation up to 7.5% of dry weight. These tubers were normal in size, some clones showed reduced tuber yield and sometimes exhibited brown sunken staining starting at tubers navel. During a storage period over of 32 weeks of one selected clone, the cyanophycin content was stable in B33-PsbY- cph A_Te tubers but the stress symptoms increased. However, all tubers were able to germinate. Nitrogen fertilization in the greenhouse led not to an increased cyanophycin yield, slightly reduced protein content, decreased starch content, and changes in the amounts of bound and free arginine and aspartate, as compared with control tubers were observed.     

Green manures and crop sequences influence alfalfa root rot and pathogen inhibitory activity among soil-borne streptomycetes

A two-year trial was conducted to determine the effects of green manures and crop sequences on plant disease, streptomycete and bacterial densities, and inhibitory activity of indigenous streptomycetes against four target pathogens. Green manure treatments, buckwheat (Fagopyrum esculentum L.), canola (Brassica napus L.), sorghum-sudangrass (Sorghum bicolor) (L.) Moench × Sorghum sudanense (Piper) Stapf.), and fallow control were tested in conjunction with three crop sequences in a Phytophthora-infested soil placed in containers. Alfalfa (Medicago sativa L.), potato (Solanum tubersoum L.), or corn (Zea mays L.) was grown in the first year, and alfalfa was grown in all containers in the second year. Compared to fallow controls, alfalfa grown in sorghum-sudangrass- or buckwheat-treated soil had significantly greater stand counts and total biomass, respectively. In addition, alfalfa grown in fallow-treated soils had the greatest Phytophthora root rot as a function of stand count. Crop rotation also had a significant effect on alfalfa root rot and yield. Potato scab disease intensity was greatest on tubers grown in fallow-treated soils, while tubers grown in canola-treated soils had the highest yields (total tuber weight). Green-manure-treated soils tended to have greater streptomycete and bacterial densities than fallow-treated soils. In addition, buckwheat- or sorghum-sudangrass-treated soils had greater proportions of streptomycetes that were antagonistic against the target pathogens than fallow-treated soils. The proportion of antagonists in soil was negatively correlated with alfalfa root rot, and positively correlated with alfalfa stand counts. Inhibitory activity of the streptomycetes was also negatively correlated with potato scab and positively correlated with potato yield. These data suggest that green manures may provide a strategy for increasing pathogen inhibitory activity within the streptomycete community in soil, and, in conjunction with crop rotation, may contribute to the control of a diverse collection of soil-borne plant pathogens on multiple crop species.     

甘肃省中部沿黄灌区轮作和连作马铃薯根际土壤真菌群落的结构性差异评估

甘肃省中部沿黄灌区是西北地区乃至全国重要的加工型马铃薯生产基地,然而因集约化种植带来的连作障碍问题已经严重影响到当地马铃薯种植业的可持续发展。采用大田试验与PCR-DGGE(Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis)技术相结合的方法,并通过真菌的18S r DNA序列分析,评估轮作(未连作)和连作条件下马铃薯根际土壤真菌群落在组成结构上的差异,以期为甘肃省中部沿黄灌区马铃薯连作的土壤障碍机理研究提供新证据。结果表明,同轮作相比,连作显著降低了马铃薯块茎产量和植株生物量,并且随着连作年限的延长,连作障碍也愈加严重。长期连作(6a)也导致马铃薯根冠比显著增加和植株收获指数的显著下降。在根际土壤真菌的种群数量和多样性上,连作和轮作间无显著差异,但在群落组成结构上差异明显。真菌18S r DNA测序分析进一步表明,马铃薯连作较轮作相比增加了Fusarium sp.和Fusarium solani以及Verticillium dahliae的种群或个体数量,而这些真菌是导致马铃薯土传病害的主要致病菌类型。根际土壤真菌群落组成结构的改变特别是与土传病害有关的致病菌滋生可能是导致当地马铃薯连作障碍的重要原因。     

Theobroxide induces tubers in potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) and flower buds in morning glory (<Emphasis Type="Italic">Pharbitis nil</Emphasis>) under non-inductive high temperatures

Induction of some plant organs including tubers and flower buds begins with sensing environmental cues, such as photoperiod and temperature in the leaves. Theobroxide has been shown to induce potato tuberization and flower-bud formation in morning glory under non-inductive photoperiodic conditions, stimulating the activity of lipoxygenase (LOX) and the synthesis of jasmonic acid (JA). In the present study, the ability of theobroxide to overcome the inhibitory effect of unfavorable high temperature on the induction of tubers in potato and flower buds in morning glory was examined. Both tuber induction and flower-bud formation under non-inductive high temperatures were promoted by the application of theobroxide at a high concentration. However, although theobroxide treatment resulted in an increase in fresh weight during potato tuber growth at 30°C, morning glory plants treated with theobroxide at 35°C failed to bloom, implying that theobroxide may assist only in flower-bud formation.     

Induction and development of potato tubers in a jar fermentor

Potato (Solanum tuberosum L.) tubers were mass propagated in a jar fermentor using a two-step method consisting of a shoot multiplication step (phase 1) followed by a tuber induction and development step (phase 2). Tuberization was observed within 2 weeks of phase 2 and the number of tubers did not increase after this culture period. In contrast, total tuber weight continuously increased for at least 10 weeks. Although the number of tubers and the total tuber weight clearly decreased under the lower temperature (17°C), this weight decrease was partially prevented by changing the temperature from 17°C to 25°C after 2 weeks of phase 2. This result indicates that tuber development can be controlled independently of induction. Lower temperatures influenced the localization and size distribution of tubers in the jar fermentor.     

Inhibition of the ADP-glucose pyrophosphorylase in transgenic potatoes leads to sugar-storing tubers and influences tuber formation and expression of tuber storage protein genes.

Transgenic potato plants were created in which the expression of ADP-glucose pyrophosphorylase (AGPase) was inhibited by introducing a chimeric gene containing the coding region of one of the subunits of the AGPase linked in an antisense orientation to the CaMV 35S promoter. Partial inhibition of the AGPase enzyme was achieved in leaves and almost complete inhibition in tubers. This resulted in the abolition of starch formation in tubers, thus proving that AGPase has a unique role in starch biosynthesis in plants. Instead up to 30% of the dry weight of the transgenic potato tubers was represented by sucrose and up to 8% by glucose. The process of tuber formation also changed, resulting in significantly more tubers both per plant and per stolon. The accumulation of soluble sugars in tubers of antisense plants resulted in a significant increase of the total tuber fresh weight, but a decrease in dry weight of tubers. There was no significant change in the RNA levels of several other starch biosynthetic enzymes, but there was a great increase in the RNA level of the major sucrose synthesizing enzyme sucrose phosphate synthase. In addition, the inhibition of starch biosynthesis was accompanied by a massive reduction in the expression of the major storage protein species of potato tubers, supporting the idea that the expression of storage protein genes is in some way connected to carbohydrate formation in sink storage tissues.     

Development of black dot disease (Colletotrichum coccodes (Wallr.) Hughes) and its effects on the growth and yield of potato plants

Seed tubers of cvs Désirée and Pentland Crown with different severities of black dot were planted in 1988 and 1989 at Rothamsted in fields in 4– or 7-course rotations, respectively. Tubers treated with prochloraz (1988) or imazalil (1989) were planted in some plots, and in others Colletotrichum coccodes inoculum was added to the soil at planting. In further experiments at Mepal, Cambridgeshire in 1989 and 1990 and at Rothamsted in 1990 on sites where potatoes had not been grown for more than 15 years, large amounts of inoculum were added to the soil around disease-free seed tubers of two (1989) or three (1990) cultivars at planting. In all experiments plants were sampled during the season and the effects of treatments on disease development, growth and yield were recorded. Disease on roots, stem bases and tubers was found early in the season and was more severe on Désirée than on Pentland Crown plants from fields in 4– or 7-course rotations. Severity increased throughout the season and with increasing amounts of disease on the seed tubers, especially with Desiree. Disease was also found on plants from disease-free tubers and was more severe in 1988 than 1989. At harvest, black dot on tubers was significantly more severe from severely affected than from disease-free seed, and was most severe where inoculum, especially large amounts, had been added at planting. Fungicide treatment decreased disease early in the season but had no effect on tuber infection at harvest. In 1989 the weight loss of seed tubers during sprouting increased with increasing amounts of black dot, but the disease had little effect on plant size through the season. At harvest the yield of ware tubers (>50 mm) decreased with severe disease but total tuber yields were not significantly affected. However, at harvest in 1988 severely affected seed yielded significantly less than healthy seed. Plants grown from mini-tubers were free from disease on sites where potatoes had not been grown for at least 15 years. Inoculum applied at planting caused severe disease on all cultivars in both years, whereas disease was slight on uninoculated plants. Inoculated plants senesced early at Mepal in 1990, but there were no significant differences in total tuber yield in any experiment. However, yields of ware tubers (>50 mm) were sometimes decreased and the total tuber number per plant increased.     

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.     

连作马铃薯根际土壤真菌种群结构及其生物效应

连作障碍已成为马铃薯产业发展的主要限制因素,为了探明马铃薯连作障碍的机理,减轻连作障碍对产量的影响,采用大田试验与PCR-DGGE分子指纹图谱技术相结合的方法,研究马铃薯连作对根际土壤真菌种群结构的影响及其生物效应.结果表明： 随着连作年限的增加,根际土壤中真菌DGGE图谱的条带数量增加,连作1～5年处理的操作分类单元(OTU)分别比对照(轮作)增加了38.5%、38.5%、30.8%、46.2%和76.9%,说明马铃薯连作使根际土壤中真菌优势种群的个体数明显增多.随着连作年限的增加,各处理间真菌种群结构的相似性越来越低.通过真菌DGGE条带的克隆测序比对发现,随着连作年限的增加,马铃薯根际土壤土传病害病原菌尖孢镰刀菌和茄病镰刀菌的数量明显增加,而球毛壳菌作为一种生防菌,连作5年时数量明显减少.连作使根际土壤中病原真菌种群过渡成为优势种群,根际微生态环境恶化,从而作用于根系,使根系活力和吸收面积下降,最终导致块茎产量大幅度-下降.     

土壤灭菌-生物有机肥联用对连作马铃薯及土壤真菌群落结构的影响

甘肃省中部沿黄灌区是全国重要的加工型马铃薯生产基地,然而因集约化生产带来的连作障碍问题已经严重影响到当地马铃薯种植业的健康发展。结合田间试验和相关的室内分析,从马铃薯块茎产量和品质、植株生理特征和土壤真菌群落结构等角度,初步评估土壤灭菌和生物有机肥联用(Ammonia Disinfection plus Bio-organic Fertilizer Regulation,ABR)对马铃薯连作障碍的防控效果。同对照相比,ABR处理的块茎产量和商品薯率分别显著增加约71.1%—152.1%和39.2%—53.3%,但块茎化学品质变化不大。ABR处理叶绿素含量和根系活力较CK均显著增加,而叶片和根系丙二醛含量显著下降。PCR-DGGE分析发现,ABR处理显著影响了马铃薯连作土壤的真菌群落结构,表现为真菌群落的多样性指数较CK相比显著下降。ABR处理还有效抑制了土传病害的滋生,植株发病率和收获后的病薯率较CK分别显著下降约67.2%—82.2%和69.1%—70.5%。采用Real-time PCR评估连作土壤中3种优势致病真菌的数量变化,显示ABR处理下立枯丝核菌、茄病镰刀菌和接骨木镰刀菌的数量在生育期内较CK均有不同程度的下降。综合来看,土壤灭菌和生物有机肥联用技术在防控甘肃省中部沿黄灌区马铃薯连作障碍上具有较大的应用潜力,而对土传病害的抑制和微生物群落结构的改善是其主要的作用机理。     

Utilization of poultry manure when cultivating potatoes in the southern steppe of the Republic of Bashkortostan

The goal of the research is to determine the effect of pretreated poultry manure and irrigation on the yield and quality of potatoes (Solanum tuberosum) grown in fields located on the southern steppe of the Republic of Bashkortostan. Field experiments were repeated every three years. During vegetation, potato leaves and tubers were studied at the full blossom, leaves’ decay, and potato harvest. The results showed that applying pretreated poultry manure at the rate of 120 t/ha favored higher tuber weight both on the rainfed and irrigated plots (from 0.23 to 0.82 kg/plant and from 0.24 to 1.02 kg/plant, respectively). On the rainfed and irrigated plots where poultry mature was not applied (control fields) the tubers’ weight ranged from 0.08 to 0.31 kg/plant and from 0.16 to 0.50 kg/plant, respectively. Upon application of 40 t/ha of manure, under irrigation, the highest marketable value of tubers was 78%. On the rainfed plots the same value was 72% when applying poultry manure at a dose of 120 t/ha. Soil moisture monitoring showed that the potatoes did not get adequate water during the growing season. When the soil moisture on irrigated plots was 70% less than its minimum water capacity, potato plantings were watered.     

The role of endophytic bacteria during seed piece decay and potato tuberization

Healthy potato tubers (Solanum tuberosum L.) cv. Kennebec were found to be internally colonized by non-pathogenic bacterial populations originating from root zone soil. These endophytic bacteria were categorized, on the basis of bioassays, as plant growth promoting (PGP), plant growth retarding (PGR) and plant growth neutral (PGN). Genera isolated from tubers included Pseudomonas, Bacillus, Xanthomonas, Agrobacterium, Actinomyces and Acinetobacter. The PGP and PGR isolates were similarly distributed throughout these genera. Bacterial populations increased in the root zone soil directly adjacent to the seed piece during and immediately following seed piece decay. Bacteria sampled at this time were capable of promoting tuber number and weight. The proportions of PGP, PGR and PGN bacteria in the root zone were altered as endophytic bacteria were released from the decaying seed piece. The study indicates that endophytic bacteria present in the seed tubers may play an important role in seed piece decay, tuberization and plant growth.     

Early generation selection for agronomic characters in a potato breeding programme

Summary A random sample of seedlings representing high, medium and poor vigour was studied for tuber colour, tuber shape, eye depth, tuber cracking, tuber yield per plant, average tuber weight and number of tubers per plant in four successive generations (F₁, F₁, F₁C₂, and F₁C₃). Based on the performance of vigour groups in various generations and inter-generation correlation coefficients, we propose a procedure for the elimination of unproductive genotypes early in the breeding programme. The data indicates that seedlings of poor vigour can be discarded at the seedling stage prior to transplantation in the field. The rejection of clones on the basis of tuber colour, tuber shape, eye depth and tuber cracking can also be initiated at the seedling stage. For tuber yield and average tuber weight a negative selection (rejection of poor phenotypes) is suggested from the first clonal generation and for number of tubers, from second clonal generation, until statistically sound replicated trials can be conducted for carrying out positive selection.