Nutrient allocation and photochemical responses of <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">canadensis</Emphasis> ‘Neva’ to nitrogen fertilization and exogenous <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> inoculation

Arbuscular mycorrhizal fungi (AMF) can promote plant growth performance, but their effectiveness varies depending on soil nitrogen (N) availability. To clarify the effectiveness of exogenous AMF along an N-fertilization gradient (0, 2, 10, 20, and 30 mM), the impacts of exogenous Rhizophagus irregularis and N on the growth, photochemical activity, and nutritional status of Populus?×?canadensis ‘Neva’ in natural soil were evaluated in a pot experiment. The results showed that the 10 mM N level was the optimal fertilization regime with the highest promotion effect on plant growth and the maximum quantum yield of photosystem II (PSII) (F_v/F_m). Excess N (20 and 30 mM) fertilization reduced the actual quantum yield of PSII (ФPSII) and the F_v/F_m of the plants. Regardless of the N availability, inoculated plants exhibited greater F_v/F_m values than did non-inoculated plants. The biomass of inoculated plants was significantly higher compared with the control under low N levels (0 and 2 mM). Under high N levels, inoculated plants showed significant increases in ФPSII. Moreover, the nutrient imbalance of plants inoculated with exogenous R. irregularis was eased by increasing P, Fe, Mn and Cu uptake in roots and higher P, Ca, Mg, Fe, Mn and Zn concentrations in leaves. Moreover, the F_v/F_m and ФPSII exhibited positive correlations with P, Ca, Mg and Zn concentrations in leaves. In conclusion, inoculation with exogenous R. irregularis can benefit plant fitness by improving the photochemical capacity and nutrient composition of poplar under different N levels.     

Phylogenetics and histology provide insight into damping-off infections of ‘Poblano’ pepper seedlings caused by <Emphasis Type="Italic">Fusarium</Emphasis> wilt in greenhouses

The ‘Poblano’ pepper crop is economically important in Mexico and throughout the world as it is used as a hot spice in food. The cultivated area of the ‘Poblano’ pepper crop is decreasing yearly for many reasons, among them a wilt disease commonly associated with Fusarium spp. This disease is a problem of field and greenhouse production plants. Moreover, it is not clear whether the pathogens that cause wilt in mature plants are the same as those involved in the damping-off symptoms and death of pepper seedlings in greenhouses. For this reason, the aim of the present study was to identify the causal agent of damping-off in pepper during seedling production, establish its relationship with the causal agent of wilting in mature plants, and determine whether histological damage in seedlings occurs. Isolates were recovered from the crown rot and stem base of 4-month-old infected ‘Poblano’ mature pepper plants and were identified using morphological and phylogenetic approaches. Fusarium oxysporum and F. solani were isolated from the crown rot and base stem, respectively. A pathogenicity test showed that both species caused damping-off in pepper seedlings. Histological studies with inoculated seedlings of both isolates showed several changes in the external cortex, epidermal cells, endodermis, Casparian strips, cell size, and xylem wall. Casparian strip rupture resulted in permeability loss and regulatory activity to maintain the cellular equilibrium inside the vascular bundles. Hence, according to these findings, producers should avoid seedling contamination by infected mature plants because the aggressiveness of Fusarium isolates can cause rapid seedling mortality.     

Roles of rootstocks and scions in aluminum-tolerance of <Emphasis Type="Italic">Citrus</Emphasis>

Four scion-rootstock combination [i.e., X/X and X/SP, ‘Xuegan’ (Citrus sinensis) grafted on ‘Xugan’ and ‘Sour pummelo’ (Citrus grandis), respectively, and SP/X and SP/SP, ‘Sour pummelo’ grafted on ‘Xuegan’ and ‘Sour pummelo’, respectively] plants were treated for 18 weeks with 0 (?Al) or 1.2 mM AlCl₃·6H₂O (+Al). Thereafter, leaf, stem and root concentrations of phosphorus and aluminum (Al), leaf and root levels of organic acids (OAs), Al-induced release of OA anions (i.e., malate and citrate), photosynthesis and chlorophyll a fluorescence (OJIP) transients were measured. Al-induced decrease of photosynthesis and damage of photosynthetic electron transport chain were less pronounced in X/X and X/SP leaves than in SP/SP and SP/X leaves, which might be related with the higher Al-induced root efflux of OA anions and leaf P concentration. C. sinensis rootstock alleviated the influences of Al-toxicity on leaf photosynthetic electron transport chain by enhancing Al-induced release of root OA anions, hence lessening Al-induced photosynthesis inhibition in SP/X plants, while the reverse was the case for C. grandis rootstock in X/SP plants. In conclusion, the tolerance of grafted Citrus plants to Al depends on the scion as well as rootstock genotype, and the scion-rootstock interaction.     

<Emphasis Type="Italic">UGP</Emphasis> gene expression and UDP-glucose pyrophosphorylase enzymatic activity in grafting annonaceous plants

Grafting is commonly used to propagate commercial fruit species to ensure that the genetic characteristics of selected clones are maintained. However, the biochemical and molecular mechanisms involved in the graft incompatibility of woody trees are not well understood. We investigated the effect of grafting in vegetative growth, UDP-glucose pyrophosphorylase expression and activity of Annonaceous grafted plants: atemoya (Annona cherimola Mill. x Annona squamosa L.) ‘Thompson’ grafted onto different rootstocks, araticum-de-terra-fria (Annona emarginata Schltdl. H. Rainer “var. terra-fria”), araticum-mirim (Annona emarginata Schltdl. H. Rainer “var. mirim”) and biribá (Annona mucosa Schltdl. H. Rainer) at different post-grafting times. The growth of atemoya grafted onto araticum-mirim was lower than that of the rootstocks araticum-de terra-fria and biribá. The results also indicated that grafting alters UGPase gene expression; showing the combination atemoya grafted onto araticum-de-terra-fria (a compatible union) the higher levels of gene expression during the early stages of grafting development. However, no significant differences were detected in UGPase enzyme activity between the graft combinations. In addition, SDS-PAGE and MALDI-TOF analyses detected similar UGPase amino acid sequences in ungrafted atemoya samples to cherimoya (Annona cherimola Mill.), a female parent of the atemoya hybrid. These findings suggest that expression of the UGPase protein is related to graft compatibility in grafted Annona plants.     

High throughput transcriptome analysis of coffee reveals prehaustorial resistance in response to <Emphasis Type="Italic">Hemileia vastatrix</Emphasis> infection

Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.     

Exogenous proline induces soluble sugar accumulation and alleviates drought stress effects on photosystem II functioning of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> leaves

The effects of exogenous applied proline (Pro), on photosystem II (PSII) photochemistry of drought stressed (DS) 4-week old Arabidopsis thaliana plants, was studied by using chlorophyll (chl) fluorescence imaging. The maximum quantum yield of PSII photochemistry (F _v /F _m) in DS plants decreased significantly to 77% of that of the control value, suggesting that DS plants could not maintain PSII function, possibly due to accelerated photoinhibition of PSII. Free Pro and total soluble sugars (SS) increased, in response to DS. Exogenous foliar application of Pro by spraying, led to a remarkable increase in the accumulation of Pro and surprisingly also of SS. Both of them served to scavenge reactive oxygen species (ROS), as it was evident by the decreased lipid peroxidation level measured as malondialdehyde (MDA). DS plants sprayed with Pro showed a tolerance to photoinhibition, this indicated by F _v/F _m being close to values typical of healthy leaves by maintaining more than 98% of PSII function. Also the higher quantum efficiency of PSII photochemistry (Φ _PSΙΙ ) and the decreased excitation pressure (1 ? q _p ) recorded for stressed leaves with Pro, lead us to conclude that Pro appears to be involved in the protection of chloroplast structures by quenching ROS. The enhanced dissipation of excess light energy of PSII, in part accounts for the observed increased resistance to DS in A. thaliana leaves with Pro. Our data pointed out that Pro signalling interacts with SS signaling pathway and provided a new insight in Pro metabolism.     

Effects of virus infection on plant growth,root development and phytohormone levels in in vitro-cultured pear plants

Virus infection is common in pear (Pyrus spp.) trees commercially cultivated in China. In this study, the growth speed, root development and phytohormone levels of virus-free and virus-infected pear cultures were comparatively investigated. Results showed that the co-infection of Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) significantly decreased the growth and proliferation of in vitro plants of P. communis cv. ‘Confenence’, but showed relatively less effect for P. pyrifolia cv. ‘Jinshui no. 2’, P. communis cv. ‘Red Clapp Favonite’ and P. sinkiangensis cv. ‘Korla’. Strong inhibition of ASGV infection to root development was observed for in vitro plants of ‘Jinshui no. 2’, but not for ‘Confenence’. Furthermore, during root induction, ASGV infection significantly increased CTKs/IAA ratio and decreased IAA/ABA ratio in the rooting region of in vitro plants of ‘Jinshui no. 2’. Nevertheless, for in vitro plants of ‘Confenence’, these values were rarely influenced by ASGV and ASPV infection. The result indicated that the changes of CTKs/IAA and IAA/ABA ratios might be responsible for rooting inhibition of in vitro plants of ‘Jinshui no. 2’.     

Host preferences and biotic potential of <Emphasis Type="Italic">Trialeurodes vaporariorum</Emphasis> and <Emphasis Type="Italic">Bemisia tabaci</Emphasis> (Hemiptera: Aleyrodidae) in tomato and pepper

Whiteflies Bemisia tabaci (Gennadius) and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) are important pests in pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.) crops in many countries. Contrary to what is observed for all other countries, in Uruguay, B. tabaci is mainly found on pepper and rarely on tomato, while T. vaporariorum is exclusively found on tomato. This study tested the oviposition preferences and biotic potential of these two whiteflies reared on both host plants. The developmental time, survival rates, longevity, fecundity and main population parameters were characterized. Both whitefly species showed different preference patterns regarding their host plants. T. vaporariorum preferred tomato instead of pepper to oviposit. Their developmental time is longer on pepper. B. tabaci preferred pepper, but the difference from tomato was not very strong. Pepper affects the biotic expression of T. vaporariorum negatively, while B. tabaci is able to develop equally on both host plants. These results show that the distribution differences of both whiteflies observed on both host plants could have a biological basis.     

Overexpression of calmodulin gene fragment from Antarctic notothenioid fish improves chilling tolerance in <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis>

Calmodulin (CaM) is a highly conserved calcium sensor protein associated with chilling tolerance in living organisms. It has four EF-hand domains for binding of four Ca²⁺, two of them located in the N-terminus, and the other two in the C-terminus. A notothenioid CaM gene fragment (CaMm), which only codes for N-terminus of CaM (with two EF-hand domains), was introduced into Nicotiana benthamiana. Effects of its overexpression on chilling tolerance in plants were explored. During 4?C or 0?C chilling treatment, both CaMm and CaM transgenic plants showed higher PSII maximum quantum yield, actual quantum yield, and soluble protein content, lower electrolyte leakage and malondialdehyde content than that of the control. The changes in these physiological indices were comparable between the CaMm and CaM transgenic plants during the treatments. These results indicate that the N-terminus of calmodulin is likely the key functional domain involved in the adaptive response to cold stress.     

Enhanced salinity stress tolerance in transgenic chilli pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) plants overexpressing the wheat antiporter (<Emphasis Type="Italic">TaNHX2</Emphasis>) gene

Transgenic chilli pepper (Capsicum annuum L.) plants tolerant to salinity stress were produced by introducing the wheat Na⁺/H⁺ antiporter gene (TaNHX2) via Agrobacterium-mediated transformation. Cotyledonary explants were infected with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBin438 that contains a wheat antiporter (TaNHX2) gene driven by the double CaMV 35S promoter and NPT II gene as a selectable marker. PCR and semiquantitative RT-PCR analysis confirmed that the TaNHX2 gene had been integrated and expressed in the T₁ generation of transgenic pepper plants as compared to the non-transformed plants. Southern blot analysis further verified the integration and presence of TaNHX2 gene in the genome of chilli pepper plants. Biochemical assays of these transgenic plants revealed enhanced levels of proline, chlorophyll, superoxide dismutase, ascorbate peroxidase, relative water content, and reduced levels of hydrogen peroxide (H₂O₂), malondialdehyde compared to wild-type plants under salt stress conditions. The present investigation clearly showed that overexpression of the TaNHX2 gene enhanced salt stress tolerance in transgenic chilli pepper plants.     

Physiological and biochemical responses of photomorphogenic tomato mutants (cv. Micro-Tom) under water withholding

In addition to mediating photomorphogenesis, phytochromes are responsible for many abiotic stress responses, acting upon biochemical and molecular mechanisms of cell signaling. In this work, we measured the physiological and biochemical responses of phytochrome-mutant plants under water stress. In tomato (Solanum lycopersicum L.), the aurea mutant (au) is phytochrome-deficient and the high-pigment-1 mutant (hp1) has exaggerated light responses. We examined the effects of water withholding on water potential, leaf gas exchange, chlorophyll fluorescence, chloroplast pigment content and antioxidant enzyme activity in au and hp1 and their wild-type cultivar Micro-Tom (MT). Initial fluorescence and potential quantum efficiency of photosystem II (PSII) photochemistry were not affected by the treatment, but effective quantum yield of PSII, electron transport rate decreased and non-photochemical quenching increased significantly in MT. Under water withholding conditions, MT had higher malondialdehyde concentration than the mutants, but au had higher activities of catalase and ascorbate peroxidase compared to the other genotypes. The tolerance of mutants to the effects of water withholding may be explained by the higher activity of antioxidant enzymes in au and by a higher concentration of antioxidant compounds, such as carotenoids, in hp1.     

Root characteristics of grafted peppers and their resistance to <Emphasis Type="Italic">Fusarium solani</Emphasis>

Root rot caused by Fusarium solani, is one of the most severe diseases in pepper (Capsicum annuum L.). Grafting has been attempted as an effective means to control the disease, but little is known about the disease resistance mechanism in grafted pepper. Therefore, we investigated the changes of biomass, cell structure, and the secondary metabolism in roots of control (non-grafted pepper) and grafted peppers using cvs. Weishi and Buyeding as rootstocks and the cv. Xinfeng 2 as a scion. After a manual inoculation, less F. solani invaded grafted pepper roots and consequently less serious injury to the root cell ultra-structure compared with the control was found. The roots of grafted pepper infected with F. solani exhibited greater biomass production and root activity than the roots of infected controls. Grafting led to an increased content of salicylic acid, benzoic acid, vanillin, lignin, and polyamines, as well as activities of phenylalanine ammonia lyase, polyphenoloxidase, and peroxidase. These results suggest that grafting improved the resistance of peppers to root rot.     

Effects of transgenic expression of <Emphasis Type="Italic">Brevibacterium linens</Emphasis> methionine gamma lyase (MGL) on accumulation of <Emphasis Type="Italic">Tylenchulus semipenetrans and key aminoacid contents</Emphasis> in <Emphasis Type="Italic">Carrizo citrange</Emphasis>

Alpha-dioxygenases (α-DOX) catalyzing the primary oxygenation of fatty acids to oxylipins were recently found in plants. Here, the biological roles of the pepper α-DOX (Ca-DOX) gene, which is strongly induced during non-host pathogen infection in chili pepper, were examined. Virus-induced gene silencing demonstrated that down-regulation of Ca-DOX enhanced susceptibility to bacterial pathogens and suppressed the hypersensitive response via the suppression of pathogenesis-related genes such as PR4, proteinase inhibitor II and lipid transfer protein (PR14). Ca-DOX-silenced pepper plants also exhibited more retarded growth with lower epidermal cell numbers and reduced cell wall thickness than control plants. To better understand regulation of Ca-DOX, transgenic Arabidopsis plants harboring the β-glucuronidase (GUS) reporter gene driven from a putative Ca-DOX promoter were generated. GUS expression was significantly induced upon avirulent pathogen infection in transgenic Arabidopsis leaves, whereas GUS induction was relatively weak upon virulent pathogen treatment. After treatment with plant hormones, early and strong GUS expression was seen after treatment of salicylic acid, whereas ethylene and methyl jasmonate treatments produced relatively weak and late GUS signals. These results will enable us to further understand the role of α-DOX, which is important in lipid metabolism, defense responses, and growth development in plants.     

Rapid identification of QTLs underlying resistance to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> in pepper (<Emphasis Type="Italic">Capsicum frutescens</Emphasis>)

Key message

Next-generation sequencing enabled a fast discovery of QTLs controlling CMV resistant in pepper. The gene CA02g19570 as a possible candidate gene of qCmr2.1 was identified for resistance to CMV in pepper.

Abstract

Cucumber mosaic virus (CMV) is one of the most important viruses infecting pepper, but the genetic basis of CMV resistance in pepper is elusive. In this study, we identified a candidate gene for CMV resistance QTL, qCmr2.1 through SLAF-seq. Segregation analysis in F₂, BC₁ and F_2:3 populations derived from a cross between two inbred lines ‘PBC688’ (CMV-resistant) and ‘G29’ (CMV-susceptible) suggested quantitative inheritance of resistance to CMV in pepper. Genome-wide comparison of SNP profiles between the CMV-resistant and CMV-susceptible bulks constructed from an F₂ population identified two QTLs, designated as qCmr2.1 on chromosome 2 and qCmr11.1 on chromosome 11 for resistance to CMV in PBC688, which were confirmed by InDel marker-based classical QTL mapping in the F₂ population. As a major QTL, joint SLAF-seq and traditional QTL analysis delimited qCmr2.1 to a 330 kb genomic region. Two pepper genes, CA02g19570 and CA02g19600, were identified in this region, which are homologous with the genes LOC104113703, LOC104248995, LOC102603934 and LOC101248357, which were predicted to encode N-like protein associated with TMV-resistant in Solanum crops. Quantitative RT-PCR revealed higher expression levels of CA02g19570 in CMV resistance genotypes. The CA02g19600 did not exhibit obvious regularity in expression patterns. Higher relative expression levels of CA02g19570 in PBC688 and F₁ were compared with those in G29 during days after inoculation. These results provide support for CA02g19570 as a possible candidate gene of qCmr2.1 for resistance to CMV in pepper.

     

Effects of drought stress on growth and chlorophyll fluorescence of Lycium ruthenicum Murr. seedlings

The present study aimed to determine effects of drought stress on Lycium ruthenicum Murr. seedlings. Our results showed that mild drought stress was beneficial to growth of L. ruthenicum seedlings. Their height, basal diameter, crown, leaf number, stem dry mass, leaf and root dry mass increased gradually when the soil water content declined from 34.7 to 21.2%. However, with further decrease of the soil water content, the growth of L. ruthenicum seedlings was limited. After 28 d of treatment, the seedlings were apparently vulnerable to drought stress, which resulted in significant leaf shedding and slow growth. However, growth was restored after rehydration. Drought treatments led to a decrease in contents of chlorophyll (Chl) a, b, and Chl (a+b) and increase in the Chl a/b ratio. After rewatering, the Chl content recovered to the content of the control plants. Under drought stress, minimal fluorescence and nonphotochemical quenching coefficient increased, thereby indicating that L. ruthenicum seedlings could protect PSII reaction centres from damage. Maximum fluorescence, maximum quantum yield, actual quantum yield of PSII photochemistry, and photochemical quenching decreased, which suggested that drought stress impacted the openness of PSII reaction centres. A comparison of these responses might help identify the drought tolerance mechanisms of L. ruthenicum. This could be the reference for the planting location and irrigation arrangements during the growing period of L. ruthenicum.     

Susceptibility of germinating cruciferous seeds to <Emphasis Type="Italic">Bagrada hilaris</Emphasis> (Hemiptera: Pentatomidae) feeding injury

Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae) is a serious pest that attacks both germinating and seedling stages of a variety of cruciferous crops grown in the Central Coast of California. B. hilaris feeding on germinating seeds can cause severe stunting and plant mortality, and little is known about the feeding preference of B. hilaris for germinating seeds of major cruciferous hosts and varieties of hosts. No-choice and choice experiments were conducted in which germinating seeds in soilless and soil settings were exposed to B. hilaris adults for 7 days. Susceptibility scores were developed using B. hilaris feeding injury sites, distorted leaves, and deformed and dead plants to determine the overall B. hilaris preference for germinating host seeds. Based on the scores, the order of preference was arugula (Eruca sativa L.)?>?turnip (B. rapa L. var. rapa)?>?mizuna (B. rapa L. nipposinica)?>?kale (B. oleracea L. acephala)?>?choi (Brassica rapa L. var. chinensis)?>?broccoli (B. oleracea var. italica Plenck)?>?cauliflower (B. oleracea L. var. botrytis)?>?lettuce (Lactuca sativa L.)?>?sweet alyssum (Lobularia maritima [L.] Desv.). The lowest feeding injury was recorded on germinating lettuce and sweet alyssum seeds. Furthermore, no-choice and choice experiments were conducted with four varieties each of arugula and mizuna, twelve varieties each of kale and choi, and nine varieties/types of leafy Asian greens. The arugula varieties ‘Wild Rocket’ and ‘Spirit’ were more damaged by B. hilaris than other varieties tested. Among mizuna varieties, ‘Beira F1’ was more attractive to B. hilaris than ‘Scarlet’ or ‘Starbor F1.’ The choi varieties ‘Tokyo Bekana,’ ‘Feng Qing Choi F1,’ ‘Joi Choi F1,’ and ‘Win-Win Choi F1’ were more attractive than ‘Rosie F1.’ The leafy Asian greens variety ‘Carlton F1’ was more attractive to B. hilaris than ‘Yukina Savon,’ ‘Tatsoi OG,’ ‘Komatsuna Summerfest F1,’ ‘Red Rain F1,’ and ‘Shungiku.’ Therefore, the results suggest that not all varieties were equally susceptible to B. hilaris feeding and possibly be utilized for further field evaluation as a trap crop or developing more resistant varieties to B. hilaris.     

A SNP-based genetic linkage map of <Emphasis Type="Italic">Capsicum baccatum</Emphasis> and its comparison to the <Emphasis Type="Italic">Capsicum annuum</Emphasis> reference physical map

Capsicum baccatum L., one of five domesticated species of Capsicum, is a valuable species in chili pepper breeding. In particular, it is a source of disease resistance against anthracnose and powdery mildew. Genetic maps and molecular markers are important to improve the efficiency of crop breeding programs. Recently, using genetic maps several researchers have identified quantitative trait loci (QTLs) for important horticultural traits and have cloned genes of interest. In this study, we constructed a genetic map of C. baccatum in an intraspecific population from a cross between ‘Golden-aji’ and ‘PI594137.’ A total of 395 high-resolution melting markers were developed based on single-nucleotide polymorphisms identified by comparing genome sequences generated through next-generation resequencing of the parents, ‘Golden-aji’ and ‘PI594137.’ The genetic linkage map contained 12 linkage groups, covered a total distance of 1056.2 cM, and had an average distance of 2.67 cM between markers. In addition, the final map was compared to the reference physical map of C. annuum ‘CM334.’ Interestingly, two major reciprocal translocations between chromosomes 3 and 5 and between chromosomes 3 and 9 were found, suggesting that these translocations might act as a genetic barrier between C. annuum and C. baccatum. Translocations between chromosomes 1 and 8 were also observed, as were previously reported in C. chinense, C. frutescens, and wild C. annuum. The synteny of other chromosomes was maintained, on the whole, except for several small inversions. The information on this genetic map will be helpful to analyze QTLs for important traits such as anthracnose resistance in C. baccatum and to study the causes of genetic barriers between C. annuum and C. baccatum.     

Adventitious shoot regeneration from in vitro leaf explants of <Emphasis Type="Italic">Fraxinus nigra</Emphasis>

In order to establish an attractive method for the production of valuable medicinal alkaloids (galanthamine and lycorine), the plants of Leucojum aestivum and L. aestivum ‘Gravety Giant’ grown in bioreactor RITA® were subjected to various concentrations of methyl jasmonate (MeJA), salicylic acid (SA), 1-aminocyclopropane-1-carboxylic acid (ACC) and 2-chloroethylphosphonic acid (ethephon) at different times of culture. The application of MeJA showed a negative effect on L. aestivum and L. aestivum ‘Gravety Giant’ plant growth. We observed that the incubation of plants during 168 h with 100 µM of MeJA resulted above two times lower F.W. (fresh weight) increments compared with control. While SA showed an inhibitory effect only on the growth of L. aestivum cultures. ACC and ethephon had a positive effect on both types of culture. Treatment with 50 µM of MeJA during 168 h stimulated galanthamine and lycorine biosynthesis in L. aestivum and L. aestivum ‘Gravety Giant’ cultures. In addition, the accumulation of galanthamine was increased when 10 µM of ACC were added to both types of culture. 10 µM of ACC stimulated also lycorine biosynthesis by L. aestivum ‘Gravety Giant’. The addition of 10 µM of ethephon had a positive effect only on lycorine production in plants of L. aestivum. SA promoted galanthamine and lycorine biosynthesis in tested plants. Indeed the highest galanthamine (0.8 mg/g dry weight: D.W.) and lycorine (1.53 mg/g D.W.) concentrations were observed in L. aestivum ‘Gravety Giant’ plants treated with 5 µM of SA during 10 h.