Effects of Tobacco Cyst Nematode on Growth of Flue-cured Tobacco

The effects of infection by tobacco cyst nematode (Globodera tabacum solanacearum) on growth of flue-cured tobacco cultivars NC 567 (resistant) and K 326 (susceptible) were evaluated in the field in 1993 and 1994. Infection by G. t. solanacearum suppressed number of leaves, plant height, and fresh weight of leaves and feeder roots. Correlations between weekly egg densities of G. t. solanacearum collected from soil and host growth during 11 weeks after transplanting (WAT) were often inconsistent between cultivars and years. However, consistent correlations were obtained between root weight and egg densities collected 9 WAT, as well as between leaf weight from susceptible K 326 and nematode egg densities 6 WAT. Leaf and feeder root weights were significantly correlated with the area under the curve for all nematodes per gram of feeder root for K 326 in 1993 and for both cultivars in 1994. Reduction in feeder root weight by G. t. solanacearum was similar for the resistant and susceptible cultivars. Reduction in fresh leaf weight by G. t. solanacearum was twice as great (P ≤ 0.07) for K 326 as for NC 567 in 1994. Incorporating nematode resistance into germplasm possessing improved yield and quality traits should produce cultivars more acceptable to growers.     

Development of Selected Tobacco Cyst Nematode Isolates on Resistant and Susceptible Cultivars of Flue-cured Tobacco

Tobacco cyst nematode (Globodera tabacum solanacearum) isolates were collected from 11 locations in Virginia, 3 in North Carolina, and 1 in Maryland. Isolates from each location were maintained and increased on flue-cured tobacco, Nicotiana tabacum cv K326. Plants of flue-cured tobacco cultivars K326 (susceptible) and NC567 (resistant) were each inoculated with 6,000 G. t. solanacearum eggs/plant. Tests were conducted over one (6 weeks) or two (14 weeks) generations of the nematode. Shoot and root weights and the number of nematodes present within a 1-g subsample of feeder roots were recorded at completion of the tests. Nematode counts were categorized by nematode life stage (vermiform, swollen, pyriform, and adult). Although significant differences in nematode development were detected among isolates, differences were not consistent across experiments. Results indicate similar virulence among G. t. solanacearum isolates on resistant and susceptible flue-cured tobacco cultivars. Therefore, plant breeders may effectively use a single G. t. solanacearum isolate when screening tobacco germplasm for resistance.     

Biological and Chemical Induction of Resistance to the Globodera tabacum solanacearum in Oriental and Flue-Cured Tobacco (Nicotiana tabacum L.)

The effects of acibenzolar-S-methyl (ASM) and four combinations of plant growth-promoting rhizobacteria (PGPR) on the reproduction of a tobacco cyst nematode, Globodera tabacum solanacearum, and growth of Nicotiana tabacum (cv. K326 and Xanthi) were tested under greenhouse and field conditions. The PGPR included combinations of Bacillus subtilis A13 with B. pumilis INR7, B. pumilis SE34, B. licheniformis IN937b, or B. amyloliquefaciens IN937a, respectively. Among the four rhizobacterial combinations, IN937a + A13 exhibited the most consistent reduction in G. t. solanacearum cysts under greenhouse and field conditions. No undesirable effects of IN937a + A13 were observed on tobacco growth under greenhouse and field conditions. Use of INR7 + A13 reduced G. t. solanacearum reproduction on flue-cured tobacco cv. K326 but not on oriental tobacco cv. Xanthi. Application of ASM reduced final numbers of G. t. solanacearum cysts, but also resulted in phytotoxicity mainly under the greenhouse conditions. When oriental tobacco seedlings were pre-grown in a IN937a + A13-treated soil-less medium, a single application of ASM at 200 mg/L one week after transplanting significantly reduced G. t. solanacearum reproduction in the field.     

Enhanced Hatching of Globodera tabacum solanacearum Juveniles by Root Exudates of Flue-cured Tobacco

Stimulation of hatching of a tobacco cyst nematode (Globodera tabacum solanacearum) by root exudates from resistant NC 567 and susceptible K 326 cultivars of flue-cured tobacco, Nicotiana tabacum, was investigated. Root exudates were collected by soaking seedlings in deionized water for 2 hours at 22 °C in the dark. Fifteen mature and uniformly sized cysts were exposed at 15, 20, or 25 °C to undiluted root exudate, root exudate diluted 1:1 or 1:3 with deionized water, or deionized water alone. Hatched juveniles were counted and removed at weekly intervals during 42 and 53 days of exposure in experiments conducted in 1994 and 1995, respectively. Root exudates from both susceptible cultivar K 326 and resistant cultivar NC 567 stimulated more hatching than deionized water at 25 °C in 1994, and at all three tested temperatures in 1995. In 1994, dilution of root exudates 1:3 reduced stimulation of hatching at 25 °C compared to undiluted exudate. Hatching at 25 °C was similarly stimulated by exposure to undiluted root exudate and exudate diluted 1:1. In 1995, both dilutions reduced stimulation of hatching by root exudates at all the temperatures.     

Control of Globodera tabacum solanacearum by Alternating Host Resistance and Nematicide

The feasibility of alternating use of resistant vs. susceptible flue-cured tobacco cultivars to improve control of Globodera tabacum subsp, solanacearum (TCN) was investigated at two Virginia locations in 1984-86. Post-harvest TCN population densities were reduced in each year of the study when fenamiphos was used with a TCN-resistant cultivar (NC 567), relative to susceptible cultivars (K 326 or Mc 944). Using NC 567 with fenamipbos also reduced preplant TCN population densities in the next growing season. Egg population densities before planting in 1986 were significantly lower in plots planted with NC 567 in 1984, even when a susceptible cultivar had been planted in 1985. Use of fenamiphos with NC 567 in 1984 and 1985 further reduced preplant egg population densities in 1986. Economic returns were significantly greater in 1984 when NC 567 was used with fenamiphos, rather than a susceptible cultivar. Treatments involving fenamiphos and (or) NC 567 in 1984 and 1985 resulted in higher economic returns in 1986 than did treatments using a susceptible cultivar without fenamiphos in both previous years. Economic returns were highest in 1986 when fenamiphos and NC 567 were used in 1984 and 1985 and a susceptible cultivar was planted in 1986.     

烤烟品种抗旱性的早期鉴定技术研究

通过固体平板培养技术,研究烟苗在干旱胁迫条件下与抗旱性密切相关的指标,对5个烤烟品种进行了抗旱性鉴定,并用隶属函数法对烤烟品种的抗旱性进行综合评价。结果表明,同一烤烟品种在干旱胁迫与正常条件下相比,其鲜重、主根长、侧根总长、根系活力、叶绿素含量及质膜透性差异显著,鲜重、侧根总长及质膜透性可作为烤烟抗旱性的敏感指标;不同烤烟品种的抗旱性有明显差异,5个品种抗旱性大小表现为:NC82 > RG11 > 红花大金元 > G28 > K326。本技术可用于烤烟品种抗性筛选。     

Temporal Efficacy of Selected Nematicides on Meloidogyne Species on Tobacco

Aldicarb, ethoprop, and fenamiphos were evaluated for their efficacy in controlling various species of root-knot nematodes on flue-cured tobacco and for their residual activity, as determined through periodic sampling and bioassays of soil taken from field plots. Field experiments were conducted at five locations over 2 years with flue-cured tobacco. Soil in plots treated with nematicides were formed into high, wide beds before transplanting with ''Coker 371-Gold'' or ''K 326'' tobacco. Residual control of Meloidogyne spp. was greatest (P ≤ 0.05) with fenamiphos (in some cases up to 10 weeks, as measured in tomato bioassays of infested soil and root fragments). Suppression of nematode reproduction by ethoprop was short-lived, and numbers of second-stage juveniles + eggs and numbers of galls in bioassays sometimes surpassed those of untreated plots within 4 weeks after treatment. Aldicarb gave intermediate control over time as compared to the other compounds. Although nematicidal efficacy of all compounds varied with site and season, fenamiphos and aldicarb generally produced the highest yields.     

Identification of Sources of Resistance to Four Species of Root-knot Nematodes in Tobacco

Resistance to the southern root-knot nematode, Meloidogyne incognita races 1 and 3, has been identified, incorporated, and deployed into commercial cultivars of tobacco, Nicotiana tabacum. Cultivars with resistance to other economically important root-knot nematode species attacking tobacco, M. arenaria, M. hapla, M. javanica, and other host-specific races of M. incognita, are not available in the United States. Twenty-eight tobacco genotypes of diverse origin and two standard cultivars, NC 2326 (susceptible) and Speight G 28 (resistant to M. incognita races 1 and 3), were screened for resistance to eight root-knot nematode populations of North Carolina origin. Based on root gall indices at 8 to 12 weeks after inoculation, all genotypes except NC 2326 and Okinawa were resistant to M. arenaria race 1, and races 1 and 3 of M. incognita. Except for slight root galling, genotypes resistant to M. arenaria race 1 responded similarly to races 1 and 3 of M. incognita. All genotypes except NC 2326, Okinawa, and Speight G 28 showed resistance to M. javanica. Okinawa, while supporting lower reproduction of M. javanica than NC 2326, was rated as moderately susceptible. Tobacco breeding lines 81-R-617A, 81-RL- 2K, SA 1213, SA 1214, SA 1223, and SA 1224 were resistant to M. arenaria race 2, and thus may be used as sources of resistance to this pathogen. No resistance to M. hapla and only moderate resistance to races 2 and 4 of M. incognita were found in any of the tobacco genotypes. Under natural field infestations of M. arenaria race 2, nematode development on resistant tobacco breeding lines 81-RL-2K, SA 1214, and SA 1215 was similar to a susceptible cultivar with some nematicide treatments; however, quantity and quality of yield were inferior compared to K 326 plus nematicides.     

Variability among Populations of Meloidogyne arenaria

Variability in reproduction and pathogenicity of 12 populations of Meloidogyne arenaria race 1 was evaluated on Florunner peanut, Centennial soybean, Rutgers tomato, G70, K326, and Mc944 tobacco, and Carolina Cayenne, Mississippi Nemaheart, and Santanka pepper. Differences among M. arenaria populations in rates of egg production 45 days after inoculation were observed for all cultivars except Santanka pepper. Differences among populations in dry top weights or fresh root weights were recorded on all cultivars. Numbers of nematode eggs produced on Florunner peanut varied from 3,419 to 11,593/g fresh root weight. On resistant tobacco cultivars (G70 and K326), one nematode population produced high numbers of eggs (12,042 and 6,499/g fresh root weight on G70 and K326, respectively), whereas the other populations produced low numbers of eggs (less than 500 eggs/g fresh root weight on both cultivars). Two variant M. arenaria race 1 populations were identified by factor analysis of reproductive rates on all nine cultivars. Differences m reproduction and pathogenicity observed among populations would affect the design of sustainable management systems for M. arenaria.     

Early season root production and zoospore infection of cultivars of flue-cured tobacco that differ in level of partial resistance to Phytophthora parasitica var. nicotianae

Root production of four cultivars of flue-cured tobacco was quantified in the field, greenhouse and phytotron. The cultivars ranged in level of partial resistance to the black shank pathogen, Phytophthora parasitica var. nicotianae, from susceptible to highly resistant. In the field, root-observation plates were installed approximately 10 cm from plants, and in greenhouse and phytotron studies, plants were grown in 4-liter containers with one sloping transparent side for root observation. Root growth was determined weekly for four weeks after transplanting in the field and daily up to 14 days after transplanting in the greenhouse and phytotron. Root tracings were made on acetate sheets placed against the sloping transparent side of the containers or against the transparent observation plates in the field following removal of soil from the outside of the observation plate. Root growth was quantified by retracing the root pattern on the acetate sheets over a digitizing tablet attached to a personal computer. Numbers of roots, root length, and mean and maximum rate of root growth were determined. Cultivars Hicks (susceptible) and K-326 (low level of resistance) had significantly larger root systems than moderately resistant G-28 or highly resistant NC 82. Differences in total root length were due to increased branching that resulted in development of significantly greater numbers of roots in Hicks and K-326. For example, between day 21 and 28, Hicks produced more than three times the number of new roots as NC 82 in the field. The mean rate of root extension observed (2.17 mm hr^–1) was similar in all four cultivars. Infection efficiency on the different cultivars was determined in the field by inoculating roots with zoospores of P. p. nicotianae. Lesions were visible as water soaked areas within 24 hr of inoculation. At 48 hr after inoculation, percentages of inoculations that resulted in lesion formation were 57, 46, 23, and 16% for Hicks, K-326, G-28 and NC 82, respectively. The possible role of rooting intensity as a mechanism of avoidance to P. p. nicotianae in tobacco cultivars is discussed.     

不同烤烟基因型钾离子通道特性研究

通过对4个不同钾营养效率的烤烟品种的根皮层细胞质膜内向跨膜钾电流进行全细胞记录,研究钾营养效率不同的烤烟基因型的钾离子通道的特性。结果表明,NC89的电流密度(pA/pF)最高,约为-80;其次是农大202及净叶黄,其电流密度分别为-60及-50;NC2326的电流密度在四个基因型中最小,大约-40。结果表明,烤烟根皮层细胞质膜跨膜内向钾电流的大小,与基因型对K+亲和力及吸收能力的大小,以及烤烟基因型钾营养效率高低有着密切的关系,可以作为一个比较可靠的参考鉴定指标,指导K+营养高效的基因型的筛选与鉴定。     

Infection,Reproduction Potential,and Root Galling by Root-knot Nematode Species and Concomitant Populations on Peanut and Tobacco

Single populations of Meloidogyne arenaria races 1 (MA1) and 2 (MA2) and M. hapla (MH), and mixed populations of MA1 + MA2 and MA1 + MH with four inoculum levels of eggs were tested on peanut cv. ''Florigiant'' and M. incognita-resistant tobacco cv. ''McNair 373'' in a greenhouse experiment. Root infection, female development, and reproduction of MA2 on peanut and MA1 on resistant tobacco were limited at 2 and 6 weeks. MA1, MH, and MA1 + MH on peanut had similar root infection (total parasitic forms per root unit) at both 2 and 6 weeks, and similar female development and reproduction potentials at 6 weeks. MA2 tended to depress root infection, female development, and reproduction of MA1 on peanut. MH had little effect on MA1 on this crop. On tobacco, MA2 population had greater incidence of root infection than did MH at 2 weeks. The two nematode species had similar development in roots at 6 weeks. All of these processes were restricted when either MA2 or MH was present together with MA1. As initial inoculum level of parasitically fit populations increased, relative infection ratio on both peanut and tobacco, and reproduction factor on peanut decreased. Populations that had high infection incidence and reproduction rates induced greater root galling than did other populations. Root galling was suppressed in the presence of antagonistic response between nematode populations.     

烟草内生细菌种群动态研究

经过对 7个田间种植的烟草品种不同栽培时期、不同组织内生细菌种群动态研究表明 ,不同品种内生细菌种群有一定程度差异。同一品种中有的内生细菌为常住菌群 ,有的为暂居菌群 ,带菌量根中最高 ,茎次之 ,叶中最低。在整个生育期中 ,7个品种内生细菌数量表现出从种子到出苗期大幅增加 ,从出苗期到十字期又大幅度下降 ,随后从缓苗期到伸根期再一次急剧增加并维持在一个较高水平。通过对烟草这一重要经济作物内生细菌种群动态变化研究 ,可为烟草病虫害生物防治和促生增产研究提供理论基础。     

Relationships Between the Population Density of Meloidogyne incognita and Growth of Tobacco

Seedlings of tobacco cultivars resistant (NC95) and susceptible (McNair 30) to Meloidogyne incognita were grown in 15-cm diameter clay pots containing steamed soil infested with 0, l, 2, 4, 8, 16, 32, and 64 eggs of M. incognita per 1.5 cm³ soil. Plants were maintained in the greenhouse for 3 weeks, and then transferred to the field for 12 weeks. Growth of tobacco was expressed separately as dry weight of leaves and as plant height. Least squares regression analysis showed that tobacco growth-nematode density interactions are in agreement with Seinhorst''s exponential model Y = m + (l-m) czp. Tobacco growth was not affected significantly as nematode density was increased from 0 to tolerance levels, which were approximately 2 and 1 eggs per 1.5 cm³ soil for the resistant and susceptible cultivars, respectively. As nematode density was increased beyond tolerance level, tobacco growth decreased sharply until a minimum yield was approached. The minimum leaf weights and plant heights of the resistant cultivar at the highest nematode density were greater than those of the susceptible cultivar.     

Genetics of Burley and Flue-Cured Tobacco Resistance to Globodera tabacum tabacum

Genotypes of burley (cultivars B-21 and B-49), flue-cured (line VA-81 and cultivar PD-4), and Connecticut broadleaf (cultivar C9) tobacco (Nicotiana tabacum) resistant (R) or susceptible (S) to the tobacco cyst nematode Globodera tabacum tabacum were crossed. F1 progeny of burley and susceptible broadleaf were selfed and backcrossed to produce additional progeny for evaluation of resistance in greenhouse experiments. Plants without adult female nematodes visible (×10 magnification) on the root surface 6 weeks after inoculation were classified as resistant, whereas those plants in which one or more females were evident were classified as susceptible. Segregation ratios for progeny of resistant and susceptible plants were not different from 3:1 and 1:1 for F2 (F1 × F1) and BC1 (F1 × S) lines, respectively, indicating that resistance in burley to G. t. tabacum is conferred by a single, dominant gene. Segregation ratios for resistance in crosses between nematode-resistant burley and flue-cured tobacco (F1 and F2 progeny) and between burley-flue-cured hybrids and broadleaf BC1 (F1 × S) and BC2 (BC1 × S) progeny were consistent with the assumption that resistance to G. t. tabacum in burley and flue-cured tobacco is conferred by the same or closely linked single, dominant gene(s).     

烘烤过程中外加淀粉类酶对烤烟淀粉降解的影响

为降低烤后烟叶中淀粉含量,研究了烘烤过程中不同外加淀粉类酶对烤烟淀粉降解的影响。结果表明：烘烤过程中,通过外加淀粉类酶来降解烤烟中的淀粉是有效的。烘烤变黄初期,不同外加淀粉类酶烟叶淀粉降解动态基本一致;变黄中期至定色前期,淀粉降解随外加酶量增加而加剧。烤后烟叶淀粉含量随外加酶量增加而减少,水溶性糖和还原糖含量随外加酶量增加而增加。方差分析表明,不同处理烤后烟叶之问淀粉含量存在极显著差异。多重比较结果表明：K326品种适宜的外加酶量为(6 60)U／g;HD的适宜外加酶量为(8 80)U／g。     

Reproduction of Meloidogyne incognita Race 3 on Flue-cured Tobacco Homozygous for Rk1 and/or Rk2 Resistance Genes

Most commercial tobacco cultivars possess the Rk1 resistance gene to races 1 and 3 of Meloidogyne incognita and race 1 of Meloidogyne arenaria, which has caused a shift in population prevalence in Virginia tobacco fields toward other species and races. A number of cultivars now also possess the Rk2 gene for root-knot resistance. Experiments were conducted in 2013 to 2014 to examine whether possessing both Rk1 and Rk2 increases resistance to a variant of M. incognita race 3 compared to either gene alone. Greenhouse trials were arranged in a completely randomized design with Coker 371-Gold (C371G; susceptible), NC 95 and SC 72 (Rk1Rk1), T-15-1-1 (Rk2Rk2), and STNCB-2-28 and NOD 8 (Rk1Rk1 and Rk2Rk2). Each plant was inoculated with 5,000 root-knot nematode eggs; data were collected 60 d postinoculation. Percent galling and numbers of egg masses and eggs were counted, the latter being used to calculate the reproductive index on each host. Despite variability, entries with both Rk1 and Rk2 conferred greater resistance to a variant of M. incognita race 3 than plants with Rk1 or Rk2 alone. Entries with Rk1 alone were successful in reducing root galling and nematode reproduction compared to the susceptible control. Entry T-15-1-1 did not reduce galling compared to the susceptible control but often suppressed reproduction.     

The negative influence of <Emphasis Type="Italic">N</Emphasis>-mediated TMV resistance on yield in tobacco: linkage drag versus pleiotropy

Resistance to tobacco mosaic virus (TMV) is controlled by the single dominant gene N in Nicotiana glutinosa L. This gene has been transferred to cultivated tobacco (N. tabacum L.) by interspecific hybridization and backcrossing, but has historically been associated with reduced yields and/or quality in flue-cured tobacco breeding materials. Past researchers have suggested the role of pleiotropy and/or linkage drag effects in this unfavorable relationship. Introduction of the cloned N gene into a TMV-susceptible tobacco genotype (cultivar ‘K326’) via plant transformation permitted investigation of the relative importance of these possibilities. On average, yield and cash return ($ ha⁻¹) of 14 transgenic NN lines of K326 were significantly higher relative to an isoline of K326 carrying N introduced via interspecific hybridization and backcrossing. The negative effects of tissue culture-induced genetic variation confounded comparisons with the TMV-susceptible cultivar, K326, however. Backcrossing the original transgenic lines to non-tissue cultured K326 removed many of these unfavorable effects, and significantly improved their performance for yield and cash return. Comparisons of the 14 corresponding transgenic NN backcross-derived lines with K326 indicated that linkage drag is the main factor contributing to reduced yields in TMV-resistant flue-cured tobacco germplasm. On average, these transgenic lines outyielded the conventionally-developed TMV-resistant K326 isoline by 427 kg ha⁻¹ (P < 0.05) and generated $1,365 ha⁻¹ more (P < 0.05). Although transgenic tobacco cultivars are currently not commercially acceptable, breeding strategies designed to reduce the amount of N. glutinosa chromatin linked to N may increase the likelihood of developing high-yielding TMV-resistant flue-cured tobacco cultivars.     

Analysis of Methylated Patterns and Quality-Related Genes in Tobacco (Nicotiana tabacum) Cultivars

Methylation-sensitive amplified polymorphism was used in this study to investigate epigenetic information of four tobacco cultivars: Yunyan 85, NC89, K326, and Yunyan 87. The DNA fragments with methylated information were cloned by reamplified PCR and sequenced. The results of Blast alignments showed that the genes with methylation information included chitinase, nitrate reductase, chloroplast DNA, mitochondrial DNA, ornithine decarboxylase, ribulose carboxylase, and promoter sequences. Homologous comparison in three cloned gene sequences (nitrate reductase, ornithine decarboxylase, and ribulose decarboxylase) indicated that geographic factors had significant influence on the whole genome methylation. Introns also contained different information in different tobacco cultivars. These findings suggest that synthetic mechanisms for tobacco aromatic components could be affected by different environmental factors leading to variation of noncoding regions in the genome, which finally results in different fragrance and taste in different tobacco cultivars.     

Interactions between Meloidogyne incognita,M. hapla,and Pratylenchus brachyurus in Tobacco

In a greenhouse pot experiment on the pathogenicity and interactions of Meloidogyne incognita, M. hapla and Pratylenchus brachyurus on four cultivars o f tobacco the cultivars ''Hicks'' and ''NC 2326'' were susceptible to each nematode and "NC 95'' and ''NC 2512'' resistant only to M. incognita.Mean heights of susceptible plants were depressed but fresh weight of tops did not differ significantly. Meloidogyne spp. increased fresh weight of susceptible (but not the resistant) roots.Reproduction of M. incognita was decreased in the presence of P. brachyurus in one case. M. hapla reproduction was less with either of the other nematodes in five out of eight cases. In 12 combinations involving P. brachyurus, reproduction of this species was depressed in seven, not affected in four and increased in one.Mechanisms involved in associative interactions were not identified but appeared to be indirect and to involve individual host-nematode responses.