General characteristics of coumarin-induced tuberization of axillary shoots ofSolanum tuberosum L. culturedin vitro

Coumarin readily stimulates tuberization of cultured axillary shoots obtained from etiolated potato sprouts. Adequate concentrations of coumarin must be continuously present in the medium to effect tuber initiation. High nitrogen, gibberellic acid, abscisic acid, indole acetic acid, or naphthalene acetic acid reduced or inhibited coumarin-induced tuberization. Gibberellic acid and high concentrations of nitrogen in medium inhibited the uptake of coumarin-3-14C by the. axillary shoot. Based on the data of physiological activities by coumarin in other plant species and our data on tuberization, we propose that coumarin may represent in part the unknown inhibitor responsible for tuber initiation with reference to the inhibitor/gibberellic acid ratio theory of tuberization.     

Effects of photoperiod on kinetin-induced tuberization of isolated potato stolons culturedin vitro

Are short-day (SD) conditions inductive or permissive for potato tuberization? This question was addressed using anin vitro culture system. Kinetin induction ofin vitro tuberization in potato stolons cultured in the dark can be partially inhibited by light treatments. Photoperiod required for a major inhibition varied with cultivars: 8 hours for cv. Red Pontiac and 16 hours for cv. Kennebec. Short photoperiods decreased kinetininduced tuberization for all cultivars. Stolons cultured on kinetin-free medium generally did not tuberize, regardless of light treatment. Effects of kinetin and photoperiod on stolon and tuber development are observed. Relationships among kinetin, photoperiod and tuber-inducing stimulus produced under short days are discussed.     

Shoot formation in potato tuber discs in tissue culture

Tuber discs of potato (Solanum tuberosum) growing on a modified Murashige and Skoog medium produced callus and embryoid bodies but failed to form shoots. However, if 0.4 ppm of 6-benzylaminopurine was added to the basal medium, shoots became visible seven weeks after inoculation.     

Optimizing the propagation of potato (Solanum tuberosum L.) by stem cuttings

Stem cuttings from potato plants are frequently used in multiplication programs for disease-free cultivars. The survival of these leafy cuttings is often very poor, and thus various techniques were investigated which were designed to increase rooting and cutting survival. Stock plants of cultivars “Caribe,” “Jemseg,” “Keswick,” “Russet Burbank,” “Sable,” “Sebago” and “Shepody” were grown in the greenhouse under long days. Promoting growth of the axillary shoots was successfully achieved by shading the shoot apex with aluminum foil. Trimming the outer leaves of the cuttings was deleterious to both survival and rooting. A rooting hormone improved root initiation and development, and the use of shade cloth or intermittent mist increased survival by reducing wilting. A rooting substrate of vermiculite: perlite (1:1) provided sufficient aeration, support and drainage for the cuttings. Antitranspirants sprayed on the stock plants prior to the excision of cuttings and also on the excised cuttings did not improve rooting or survival. The addition of activated charcoal (1% w:w) increased rooting of the cuttings and improved vigor of potted plants obtained from the cuttings.     

油菜玻璃腋生枝的形态特征及影响其形成的因素

切除顶芽和叶片的油菜带节枝段,在含BA1mg/l和NAA0.1mg/l的MS培养基上,其腋生分生组织通常发育成三种类型腋生枝:正常的、半透明的或玻璃化的。顶芽和叶片的存在抑制玻璃腋生枝形成,叶片对腋生分生组织的正常发育也具有一定的作用。玻璃腋生枝的形成与培养基中的细胞分裂素有关。腋生分生组织发育成一定大小的正常腋芽后,不能形成玻璃腋生枝。     

Eradication of potato virus X from potato by ribavirin treatment of cultured potato shoot tips

Ribavirin treatment of cultured potato shoot tips was tested as a means of eradicating potato virus X (PVX) from two potato cultivars. Shoot tips were cultured on liquid medium containing 0, 1, 10 or 100μg/ml ribavirin. Cultures were evaluated periodically for viability, and scored for vigor on a relative growth scale. Developed plantlets were assayed for PVX by transmission tests toGomphrena globosa. Ribavirin treatment was phytotoxic at all tested concentrations, and lethal to all cultivars treated at 100 μg/ml. Treatment also delayed plantlet development by up to 2 months at 1 and 10 μg/ml as compared with nontreated controls. PVX assays indicated that 80 and 83% of the plantlets were free of PVX following treatment with 10 μg/ml for cultivars Russet Burbank and Red McClure, respectively. Five and 6% of the plantlets developed from the 1 μg/ml treatment were PVX-free, whereas 0 and 2% of the controls were PVX-free for the same cultivars. Six to 8 months were required to develop plants from shoot-tip cultures treated with 10 μg/ml ribavirin.     

Detection of potato viruses X and S in tissue culture plantlets

The latex agglutination test (LAT) and enzyme-linked immunosorbent assay (ELISA) were evaluated in separate studies for their ability to detect potato virus X (PVX) and potato virus S (PVS) in tissue culture plantlets. Healthy and infected clonal lines of several potato cultivars were used. LAT was unsatisfactory because only low levels of agglutination were obtained with infected samples, and because variable and inconsistent results were obtained with both healthy and infected clones. ELISA, however, consistently gave high spectrophotometric readings and intense visual reactions for infected but not for healthy clones. The results indicate that ELISA can be used to detect PVX and PVS in tissue culture plantlets, and in programs where tissue culture is employed, early detection and elimination of infected plantlets is possible.     

Effects of soil fumigation with Telone C on soil and plant nitrogen,verticillium wilt and yield of Norland and Norgold potatoes

Two fields in Weld County, Colorado were fumigated in the fall or spring with Telone C in an attempt to restore their productivity. The soils were Julesburg sandy loams and had been cropped to either corn or potatoes for many years. Observations were made over three years on the following: soil ammonium and nitrate nitrogen carry over, seasonal changes in these two forms of nitrogen, petiole nitrate concentration,Verticillium population, verticillium wilt expression,Verticillium stem infection and potato yields of two varieties. No significant differences occurred between control and fumigated blocks in the carry over of soil ammonium or nitrate nitrogen. During the growing season of the first year, no significant differences in soil nitrate concentrations occurred, while during the second and third year at certain times, all fumigated blocks contained significantly less nitrate nitrogen than the control block. Significant seasonal differences measured between blocks in soil ammonium concentration were not related to fumigation. Petiole nitrate concentrations while similar during the first year, were significantly higher in most fumigated treatments the second year and generally lower than the controls in the third year. Verticillium populations were not consistently affected by fumigation. Fluctuations not associated with fumigation prevented drawing definite conclusions. Both spring and fall fumigation reduced wilt severity in the two varieties. Effects of spring fumigation carried through two seasons while fall fumigation did not. Plant infection, determined by culturing stem sections, corresponded with the wilt severity ratings. Fall fumigation increased market yields of Norland potatoes but the increase was not statistically significant. Usually, both fall and spring fumigation significantly increased market yield of Norgold potatoes and over three years, spring fumigation was significantly better. Yield increases resulting from fumigation were considered greater than could be explained from alteration in the soil ammonium to nitrate ratios and/or verticillium wilt control as measured in this study.     

Efficient shoot regeneration from direct apical meristem tissue to produce virus-free purple passion fruit plants

Purple passion fruit is an important fresh table fruit. At present, the production of passion fruit is decreasing because of the spread of viral diseases throughout the planting area. The aim of this research was to propagate virus-free plants using a tissue culture technique involving the apical meristem of purple passion fruit. Shoot tips were excised to a length of 2 mm and the shoots were regenerated by culturing on Murashige and Skoog medium supplemented with different concentrations of benzyladenine (BA) (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/l) and 1-naphthaleneacetic acid (NAA) (0.2, 0.5, 1.0, 1.5 and 2.0 mg/l). Root formation was promoted using different concentrations of indole-3-butyric acid (IBA) (0.0, 0.2, 0.4, 0.6, 0.8 and 1.2 mg/l). A greater number of shoots were produced with BA concentrations of 1.0 and 1.5 mg/l than with any other BA concentrations tested (less than 1.0 mg/l or greater than 1.5 mg/l). However, when NAA at any concentration was included in the medium, no shoots were produced in culture. The cultures including 1.0 mg/l and 1.5 mg/l BA were then subcultured four times every two weeks. Initially, the tissue cultured in the 1.5 mg/l BA medium grew faster than that cultured in the 1.0 mg/l BA medium. The tissue cultured with 1.5 mg/l BA generated many short shoots, whereas the tissue cultured with 1.0 mg/l BA, generated long shoots that could be subcultured into individual plants. These regenerated shoots were assayed for the presence of the passion fruit woodiness virus using ELISA or a test strip kit; only virus-free shoots were used for further propagation. Root formation was very good in IBA concentrations of 0.4 and 0.6 mg/l. Thus, virus-free plants could be successfully regenerated directly from the apical meristem.     

Maintenance,symptoms and distribution of potato viruses X,S, Y,A, M and leafroll in potato tissue culture plantlets

Maintaining potato viruses X, S, Y, A, M and leafroll in tissue culture plantlets is a convenient, cost and space effective alternative to the use of greenhouse plants. Of these six viruses, only certain strains of PVX induced symptoms in tissue culture plantlets. Nevertheless, all infected tissue culture plants were found to be more reliable than greenhouse grown plants as virus-infected controls in enzyme-linked immunosorbent assay (ELISA) testing. Another advantage of maintaining viruses in tissue culture plantlets was the elimination of contamination by other viruses or other pathogens. Leaves, stems, and roots of virus infected plantlets were tested separately for antigen levels by ELISA. In these tests, the stems and leaves of all but PVA infected tissue culture plants consistently gave positive ELISA values. In contrast, root tissue from PVY infected tissue culture plantlets was not reliable for PVY detection. In all cases, the viruses detected in the original source material were detected in the resulting tissue culture plantlets.     

Plant regeneration from tuber discs of potato (Solanum tuberosum L.) using 6-benzylaminopurine (BAP)

Summary Shoots, roots and callus were formed from tuber discs of potato, cultivar Désirée, when grown in vitro on the basal medium of Murashige & Skoog (1962) (MS) supplemented with 2,4-D and/or BAP. Callus was formed in MS medium with 1 mg l⁻¹ BAP plus 0.5 mg l⁻¹ 2,4-D, callus and roots were formed in MS with 1 mg l⁻¹ BAP plus more than 0.5 mg l⁻¹ 2,4-D and shoots were formed directly on tuber discs cultured on MS medium with 1 mg l⁻¹ BAP without the addition of 2,4-D. Nodules produced at the explant surface after the 4th week increased in size following subculture onto the same medium (MS+BAP alone), and 2 to 6 shoots developed from each nodule. After 9 weeks total time in culture, these shoots were excised and transferred as cuttings to MS medium without growth regulators, after which roots developed and plantlets were formed. A histological study of the explants at the sites of nodule formation indicated that the shoots developed from meristematic zones initiated within small outgrowths of tissue similar to those occuring in adventive organogenesis but the presence of shoot and root meristems associated with the same axis suggests the formation of somatic embryos.     

Eradication of potato virus Y and S from potato by chemotherapy of cultured axillary bud tips

Ribavirin (virazole) treatment of cultured axillary bud tips (3-4 mm) was tested as a method of eradicating potato virus Y (PVY) and potato virus S (PVS) from two potato cultivars, Norchip and Desiree. Ribavirin treatment was phytotoxic at all concentrations tested, but cultivars treated at 5 mg/1 were visually similar to the nontreated control cultures after 20 weeks. The buds treated with ribavirin at 20 mg/1 had a survival rate of only 30–40%. Virus assays indicated that 2,74,82, and 89% of the plants were free of PVY, and 1, 71, 83 and 90% were free of PVS, 20 weeks following treatment with 0, 5, 10, and 20 mg/1, respectively. Virus assays indicated that 2, 74, 82, 89% of the plants were free of PVY, and 1, 71, 83 and 90% were free of PVS, 20 weeks following treatment with 0, 5, 10, and 20 mg/1 ribavirin treatment respectively for cultivar Norchip. Desiree cultivar assayed 2, 69, 80 and 86% free of PVY and 2, 74, 85 and 93% free of PVS, 20 weeks following treatment with 0, 5, 10, and 20 mg/1 ribavirin treatment respectively.     

Response of selectedSolanum species to virus eradication therapy

Thirty-fourSolanum genotypes infected with one or more potato viruses were evaluated in anin vitro virus eradication system combining heat therapy and the antiviral chemical ribavirin. This system was compared within vitro heat therapy or ribavirin treatment alone. The effect of expiant size on virus eradication following therapy was also evaluated. Infected plantlets were established in ribavirin lacking and amended medium; half of the plantlets in each medium were subjected to a 30-day heat treatment (35 C/31 C, 4-hr alternating periods), while the remaining plantlets were kept at room temperature (23 C). Following therapy, plantlets were tested quantitatively (OD at A₄₀₅) for the appropriate virus by enzyme linked immunosorbent assay (ELISA) and divided into 3 categories: low(< -0.05 OD), medium (> 0.05 to < 1.0 OD) and high (> -1.0 OD). Three plantlets were selected per group and further propagated as nodal cuttings or meristemtips. Regenerants which tested negative (< -0.05 OD) were transplanted to plug trays in a greenhouse and were retested after a 30-day grow-out period. Seventy-four and 91% of the genotypes yielded virus-free (VF) plants following propagation as nodal cuttings and meristem-tips, respectively. The combined therapies generated twice as many VF plants as thermotherapy alone and 5% and 15% more VF plants for nodal cutting and meristemtip derived plantlets, respectively, than for chemotherapy alone. Eleven of the 34 genotypes exhibited susceptibility to heat, but VF plants were obtained from survivors or by shortening the thermotherapy period. The efficiency of the system was enhanced by selecting plantlets for propagation which tested < -0.05 OD after treatment with 92%, 93%, 71%, and 100% of them remaining VF from potato viruses S, X, Y and leafroll, respectively. Twice as many VF plantlets were produced when treated plantlets were then propagated from meristem-tips than from nodal cuttings, but the rapid regeneration rate of plantlets from nodal cuttings makes it an attractive propagation method. Plantlet evaluation by ELISA after therapy to select plantlets with a high probability of producing VF plantlets and after further propagation to assure virus freedom are strongly recommended.     

Phytophthora erythroseptica in Peru: Its identification and pathogenesis

EightPhytophthora isolates cultured from diseased potato plants and tubers in Peru were identified asPhytophthora erythroseptica Pethyb. var.erythroseptica. This pathogen was not previously reported from South America. The isolates were pathogenic to potato plants but they differed in virulence. Injuring roots prior to inoculation enhanced infection and disease development but all isolates infected noninjured roots. Continuous high soil moisture during incubation favored disease development. Under these conditions, disease developed at soil temperatures from 10 C to 30 C, but most rapidly at 25 C and 30 C. Plants grown 60 days before inoculation developed symptoms earlier and more rapidly than plants grown 15-, 30-, and 45 days. Four concentrations of mycelial-suspension inocula did not alter disease development, but zoospore inoculum induced symptoms and plant death more rapidly than mycelial suspensions at 25 C.     

Protein,starch, and non-protein nitrogen accumulation in high protein hybrids and low protein cultivars

Tuber protein, starch, non-protein nitrogen and percent dry matter content were measured during tuber development of four Phureja-Tuberosum-Andigena hybrids and two Tuberosum cultivars. Percentages of starch and protein in the tuber dry matter were curvilinearly related to tuber size in all six clones. The amounts of starch and protein in a tuber were linear functions of tuber size. Rates of protein accumulation in some of the hybrids were higher than those of either cultivar. Starch accumulation rates were similar among all six clones. The high protein character of at least one of the hybrids was associated with starch containing storage tissue. Differences between high and low protein clones were mainly related to quantitative differences in the subunit composition of the protein. The ratio of non-protein nitrogen to protein N was similar among the six clones. Relationships between percent dry matter and starch content were not consistent among the six clones.     

Potato minitubers: Technology validation in Mexico

The effects of four previously reportedin vitro tuber-inducing modified culture media on the potato cultivars Atzimba and Juanita were studied under three illumination regimes. Tuberization from stem segments started 7–18 days after transfer for all the cultivar-light-medium combinations, but this early start was not related to tuber yield. Atzimba tuberized more than Juanita (1.0 and 0.8 tubers/plant respectively), and 8 h light was optimal. The medium by Wang Hu (WH) promoted the greatest tuberization for cultivar Atzimba (1,198 mg/plant) and that by Palmer-Smith (PS) for Juanita (290 mg/plant). The modified Stallknecht’s (ST), on the contrary, slightly stimulated tuberization under all illumination conditions. The presence of cytokinins and light favored the tuberization process. Dormancy was not observed in any of the microtubers, regardless of the tuber-inducing treatment.     

Kinetin,thermotherapy, and tissue culture to eliminate potato virus (PVX) in potato

PVX infected plantlets from two potato cultivars grownin vitro with 0.3, 3, 30 or 300 ppm kinetin were exposed to temperatures of 28 or 35 C. After 3 wk, axillary buds were isolated and grown aseptically in organogenic media, followed by PVX testing by ELISA. The serological test was also run on whole plantlets at the end of the kinetin-temperature exposure. No donor plants exposed to 28 C nor the plantlets derived from their buds gave an ELISA (-) reaction, regardless of the kinetin content of the media or that of the cultivar. At 35 C the virus was suppressed to undetectable levels in several whole plantlets. In the cultivar Alpha, 2 out of 6 resulting plantlets after isolation of buds were virus-free in the presence of 3.0 mg/1 kinetin during the treatments. From Atzimba, about 15–40% of the regenerated plants were ELISA (-), without any relationship to the cytokinin content in the media. Heat had a stronger influence on virus elimination than kinetin and the results varied with the cultivar.     

Breeding behavior of resistance to green peach aphid in tuber-bearing Solanum germplasm

Fifty-seven tuber-bearingSolanum progenies including species and interspecific hybrid derivatives were evaluated for resistance to green peach aphid (GPA),Myzus persicae (Sulzer), using an excised leaflet test. The progenies segregated for resistance and most contained some highly resistant clones. Resistance appeared to be partially dominant and genotype-environment interaction was low. Heritability estimates of 50–60 percent were obtained from both diploid and tetraploid populations.     

Plant growth response from aged potato seed-tubers as affected by meristem selection and NAA

Plant growth from Russet Burbank seed-tubers of different ages was compared as affected by source of meristematic tissue (apical or basal) and NAA treatment. In controlled environment studies, single-eye seedcores from 17-mo-old seed-tubers produced twice as many shoots compared to those from 5-and 29-mo-old seed-tubers. Seed-tuber age and shoot dry weight/core were inversely related. Dry weight/shoot decreased as age increased to 17 mo and then remained unchanged from 17 to 29 mo. Apical meristems lost growth potential faster than basal meristems as age advanced. Meristem physiological age apparently depends on relative position of the buds as well as on the chronological age of the seed-tuber. NAA (10–40 mg/l) decreased the number of shoots/core, had no effect on shoot dry weight/core but increased the dry weight/shoot when applied to seedcores from 20-mo-old seed-tubers. On seedcores from 8-mo-old seed-tubers. NAA decreased the shoot dry weight (g/core, g/shoot). In field studies, NAA (100 mg/l) increased the total tuber yield of plants grown from seedpieces cut from 8-and 20-mo-old seed-tubers. These increases were manifested in age-specific effects on tuber grade. The effects of seedtuber age on plant growth and tuber yield were partially overcome by seedpiece treatment with NAA.     

Petiole nitrate content of Maine-grown Russet Burbank and Shepody potatoes in response to varying nitrogen rate

Russet Burbank and Shepody potatoes were grown with at-planting nitrogen fertilizer rates varying from 0 to 270 kg ha^?1 following small grains and red clover. Petiole samples were collected from the 4th and 5th leaflets at four to six dates each during the 1986 to 1989 growing seasons. The samples were dried and analyzed for NO ₃ ^? N content. Petiole NO ₃ ^? N levels were strongly related to nitrogen rate regardless of cultivar, growing season, and cropping system. Differences among treatments and NO ₃ ^? N content varied substantially as the growing season progressed with petiole NO ₃ ^? N levels declining rapidly in underfertilized plots as the plants aged. Petiole NO ₃ ^? N levels were higher at midseason following red clover than following small grains. Sampling 50 to 60 days after planting (DAP) is most appropriate as a tool for scheduling supplemental nitrogen applications. At 50 DAP, critical petiole NO ₃ ^? N levels were 1.6 and 1.7% for Russet Burbank and Shepody, respectively. Petiole NO ₃ ^? N levels above 2.2% at 50 DAP resulted in lower yields of Russet Burbank than when petiole NO ₃ ^? N levels were in the 1.6 to 2.2% range. Petiole NO ₃ ^? N testing should be particularly useful as a diagnostic tool in management strategies which make maximum use of previous crop residues, organic amendments, and soil reserves as nitrogen sources.