Chloride and sodium excluding capacities of citrus rootstock germplasm introduced to Australia from the People's Republic of China

The responses to root zone salinity (0, 25 and 50 mM NaCl) by 40 citrus rootstock genotypes introduced from the People's Republic of China, measured as shoot chloride (Cl⁻) and sodium (Na⁺) ion accumulation, growth and dry matter accumulation, were investigated under glasshouse conditions. Two experiments, one using aerated nutrient solutions and the other irrigated sand cultures, were conducted with plants grown from rooted cuttings taken from representative trees of different mandarin (Citrus reticulata Blanco and C. erythrosa Hort. ex Tan.), yuzu (C. junos Sieb. ex Tan.), Ichang papeda (C. ichangensis Swing.), sour orange (C. aurantium L.) and trifoliate orange (Poncirus trifoliata (L.) Raf.) genotypes. Two standard genotypes, viz. Rangpur lime (Citrus x limonia Osbeck.) and William's trifoliate orange were included in the experiments as reference controls.     

Evaluation of strains of Poncirus trifoliata and trifoliate orange hybrids for resistance to Phytophthora root rot

Ten strains of Poncirus trifoliata (L.) Raf. and 6 trifoliate orange hybrids were evaluated for resistance to root rot caused by Phytophthora nicotianae B. de Haan var. parasitica (Dast.) Waterh. A high degree of resistance was shown by Rubidoux trifoliate and Trifoliate orange (Srirampur). Trifoliate orange hybrids, in general, were less resistant than P. trifoliata strains. The usefulness of different parameters in the assessment of root damage are briefly discussed.     

Transformation of trifoliate orange with rice chitinase gene and the use of the transformed plant as a rootstock

RCC2, cDNA clone encoding rice class-I chitinase, was introduced into trifoliate orange (Poncirus trifoliata Raf.). Chitinase activity of the transformed plants was higher than that of the non-transformed plants. The introduced gene was not detected in the scions grafted onto the transformed plants. The effect of the introduction of foreign gene into rootstocks on the scions was discussed.     

Agrobacterium tumefaciens-mediated genetic transformation and plant regeneration from a complex tetraploid hybrid citrus rootstock

Agrobacterium-mediated genetic transformation of a tetraploid “tetrazyg” citrus rootstock selection ‘Orange #16’ [Nova mandarin (Citrus reticulata Blanco) + Hirado Buntan pummelo (Citrus grandis L. Osbeck)] × [Cleopatra mandarin (C. reticulata Blanco) + Argentine trifoliate orange (Poncirus trifoliata (L.) Raf.)] was performed. Juvenile epicotyl segments were transformed with a construct containing a bifunctional egfp–nptII fusion gene under the control of an enhanced double CaMV 35S promoter. Our protocol resulted in a reasonable transformation efficiency of 18%. Stable integration of the transgene was confirmed by visual observation of EGFP expression, PCR and Southern blot hybridization. The purpose of this work was to investigate the amenability of novel citrus rootstock germplasm being developed for improved tree size control, soil adaptation, and disease resistance, to existing transformation technologies. Seed trees of such transgenic tetraploids also have potential as trap plants containing potent insecticidal transgenes, due to their inedible fruit and inherent crossing barriers with conventional commercial diploid scion cultivars, and could be planted around producing citrus groves.     

Production and evaluation of transgenic sweet orange (Citrus sinensis Osbeck) containing bivalent antibacterial peptide genes (Shiva A and Cecropin B) via a novel Agrobacterium-mediated transformation of mature axillary buds

Transgenic plants of Jincheng orange (Citrus sinensis Osbeck) and Newhall navel orange (C. sinensis Osbeck) containing antibacterial peptide genes Shiva A and Cecropin B were successfully obtained by a novel Agrobacterium tumefaciens-mediated transformation of the mature axillary buds. PCR and Southern blot analysis of the transgenic plants verified that the Shiva A and Cecropin B genes were integrated into the citrus genome. The transgenic plants began to blossom and bear fruit in the 2nd year after grafting on trifoliate orange (Poncirus trifoliata Raf) rootstock in greenhouse. Water-soluble extracts from transgenic citrus leaves exhibit in vitro suppressive effects on Xanthomonas axonopodis pv. citri, suggesting that the expressed products of Shiva A and Cecropin B in citrus retain their native antibacterial activities. Artificial inoculation in greenhouse and open field further indicates significantly increased resistance of transgenic plants to X. axonopodis pv. citri when compared with non-transgenic lines. No significant difference was found in the content of total soluble solids, total acidity, reduced sugar content and other fruit characteristics between transgenic and non-transgenic plants. In this present study, 11 transgenic lines were obtained from 40 transgenic lines, showing enhanced resistance to citrus canker disease.     

Horticultural performance of ‘Folha Murcha’ sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. ‘Folha Murcha’ sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, São Paulo State, Brazil, to evaluate the horticultural performance of ‘Folha Murcha’ sweet orange budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid ‘Rangpur’ lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’, and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the ‘Sun Chu Sha Kat’ mandarin (C. reticulata Blanco); the ‘Sunki’ mandarin (Citrus sunki (Hayata) Hort. ex. Tanaka); the ‘Rangpur’ limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (C. sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 7 m × 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the ‘Flying Dragon’ trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto ‘Cravo FCAV’ ‘Rangpur’ lime. Both ‘Rangpur’ lime rootstocks and the ‘Sunki’ mandarin induced higher tree tolerance to drought. The ‘Flying Dragon’ trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to ‘Folha Murcha’ trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on ‘Folha Murcha’ tree performance. Among the 12 evaluated rootstocks, the ‘Flying Dragon’ trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of ‘Folha Murcha’ trees, and may be better suited for high-density plantings.     

Effect of arbuscular mycorrhizal fungal inoculation on root system architecture of trifoliate orange (Poncirus trifoliata L. Raf.) seedlings

Plant root system architecture is essential characteristics in relation to nutrient acquisition by root system from soil volume. Many environmental factors can affect the establishment of root system architecture, e.g. arbuscular mycorrhizal (AM) fungi. We inoculated the trifoliate orange (Poncirus trifoliata L. Raf.) seedlings with four AM fungal species in rhizoboxes, with non-inoculated seedlings as control. Using the WinRHIZO^® image analysis system, the root system architecture of seedlings was characterized. Results indicated that AM colonization did not affect the tap root length, the average root diameter, the basal root growth angle in spite that four AM fungal species exerted differential influence on the plant growth. Contrastingly, AM colonization significantly reduced the total root length, the root volume, the root surface area, but promoted the formation of lateral roots of high order. In addition, AM colonization induced more fine roots and less coarse roots. To our knowledge, it is the first report on the influence of AM fungi on the distribution of root diameter size classes. The mechanisms and implication of AM fungi on root system architecture is discussed.     

Arbuscular mycorrhizal fungi alleviate iron deficient chlorosis in Poncirus trifoliata L. Raf under calcium bicarbonate stress

Summary

The effect of the arbuscular mycorrhizal (AM) fungus Glomus versiforme on chlorosis due to iron deficiency in trifoliate orange (Poncirus trifoliata L. Raf) was studied under calcium bicarbonate stress in sand culture. Seeds were sown in a mixed substrate [1:1 (v/v), perlite:sand] with or without mycorrhizal inoculum. The test was carried out with four treatments: Hoagland and Arnon nutrient solution with normal, no added iron, or 50% normal iron at two levels of calcium bicarbonate. The results showed that the proportion of the total root length infected by the AM fungus was ≥ 83%, and was depressed under Fe deficiency. No root colonisation was found with no added AM fungus (NM plants).

Colonisation by AM fungus resulted in higher dry weights of both shoots and roots compared to NM treatments, suggesting that the AM fungus accelerated plant growth. The activities of peroxidase and catalase in NM plants were lower than in AM fungus-inoculated plants. The results indicate that AM fungus alleviated calcium bicarbonate stress on the cell membranes of host plants. The results also showed that AM fungus increased chlorophyll concentrations, iron contents, and the Fe:P ratios of whole plants, and decreased 50(10P+K):Fe ratios, which implied that the uptake and translocation of iron was strengthened. Therefore, AM fungus had a positive effect on ameliorating chlorosis due to iron deficiency in P. trifoliata L. Raf.     

Differences of hyphal and soil phosphatase activities in drought-stressed mycorrhizal trifoliate orange (Poncirus trifoliata) seedlings

Differences of hyphal and soil phosphatase activities between mycorrhizal and non-mycorrhizal plants were less studied under drought-stressed (DS) conditions. In a pot experiment, fungal alkaline phosphatase (FALP), and succinate dehydrogenase (FSDH), soil phosphatase activity, both soil and plant P contents were compared in 6.5-month-old trifoliate orange (Poncirus trifoliata) seedlings under 80 days of DS with or without inoculations by arbuscular mycorrhizal fungi (AMF, Glomus diaphanum, Glomus mosseae or Glomus versiforme). Plant growth and biomass production under DS were significantly higher in mycorrhizal than in non-mycorrhizal seedlings. Both the FALP and the FSDH activities under DS were significantly reduced in these three Glomus inoculated seedlings. In general, similar soil neutral and alkaline phosphatase activities, but significantly higher soil acid and total phosphatase activities, were exhibited in mycorrhizal than in non-mycorrhizal seedlings under both the well-watered (WW) and the DS. Both leaf and root P contents were significantly higher in the AM colonized seedlings, but soil available P contents were lower in the growth media with AM seedlings. Our results showed that higher hyphal enzymes’ activities, soil acid and total phosphatase activities, and plant P contents in AM colonized seedlings, particularly in Glomus mosseae-colonized seedlings and/or under DS, would result in a better growth of the host plants, which might be the basis for enhancing drought tolerance in plants.     

Somatic hybridization of high yield,cold-hardy and disease resistant parents for citrus rootstock improvement

Summary

Increasing losses of trees to diseases, expansion of plantings into more marginal production areas and the need to control tree size to reduce harvesting costs have elevated the demand for new improved citrus rootstocks in Florida. A major strategy of the CREC rootstock improvement programme has been to use protoplast fusion to produce allotetraploid somatic hybrids that combine complementary rootstock germplasm. Tetraploid citrus rootstocks have been shown to have a built-in tree size control component due to polyploidy. This report focuses on the incorporation of newly identified superior rootstock germplasm into the somatic hybridization programme. Poncirus trifoliata 50–7, a selection of trifoliate orange selected for superior resistance to Phytophthora nicotianae, was hybridized with sour orange, Changsha mandarin, Navel orange, and a seedy white ‘Duncan’ type grapefruit. High-yielding Benton citrange (Citrus sinensis 3 Poncirus trifoliata) was hybridized with Changsha mandarin and sour orange. More than 200 plants of each of these six new somatic hybrids were propagated via tissue culture and rooted cuttings. To assess their horticultural performance, all the somatic hybrids were budded with commercially important scions and planted in replicated trials representing the two most important soil types in Florida. Seed trees of each somatic hybrid have also been planted in the field to determine fruiting potential and level of polyembryony.     

The effects of low boron on growth,gas exchange,boron concentration and distribution of ‘Newhall’ navel orange (Citrus sinensis Osb.) plants grafted on two rootstocks

The effects of low boron (B) on plant growth, photosynthesis, B concentration and distribution of ‘Newhall’ orange (Citrus sinensis Osb.) plants grafted on either Trifoliate orange (Poncirus trifoliata (L.) Raf.) or Carrizo citrange [C. sinensis (L.) Osb. × Poncirus trifoliata (L.) Raf.] rootstocks were investigated. One-year-old plants of the two scion-rootstock combinations were grown for 183 days in sand:perlite (1:1, v/v) medium under greenhouse conditions. The plants were irrigated with half-strength Hoagland's nutrient solutions containing four B concentrations (0.01, 0.05, 0.10 and 0.25 mg l⁻¹). The growth of root, stem of scion and leaves was less affected by low B treatments when ‘Newhall’ scion was grafted on Carrizo citrange than on Trifoliate orange. Thus, the growth of scions under low B conditions was mainly depended on the rootstock used, i.e., Carrizo citrange-grafted plants were more tolerant to low B compared to the plants grafted on Trifoliate orange. Boron concentrations in all plant parts decreased significantly by decreasing the B supply in the nutrient solution. Leaves were the dominant sites of B accumulation and showed the greatest reduction in B concentration compared to the other plant parts, as B concentration in the nutrient solution decreased. Irrespective of the rootstock, B levels in the upper–younger leaves were substantially higher than in basal-older leaves when plants were exposed to low B concentrations (≤0.05 mg l⁻¹), suggesting that under such conditions B was preferentially translocated to upper–younger leaves to support their growth. Furthermore, B distribution in different plant parts indicated that there was a restriction in translocation of B from root to scion tissues (stems and leaves of scion) under conditions of limited B availability. In addition, low B treatments decreased leaf photosynthetic rate, stomatal conductance and transpiration rate but increased intercellular CO₂ concentration in the leaves of ‘Newhall’ plants, irrespective of the rootstock used.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.     

沙田柚在不同枳选系砧木上的早期表现

比较了4个枳选系作沙田柚砧木的早期表现。结果表明,以不同枳选系作砧的沙田柚在树冠生长量、早结性等方面有差异。以小叶小花类型枳作砧木的,树冠小,结果早,以小叶大花类型枳作砧木的,树冠大小中等,但结果较迟。     

Evaluation of rootstocks for the Cyprus local lemon variety ‘Lapithkiotiki’

The effect of various rootstocks on yield, yield efficiency, tree size and fruit quality of the local lemon variety ‘Lapithkiotiki’ (Citrus limon (L.) Burm. F.) was studied under Cyprus conditions. Total cumulative yield over 13 years of production was the highest on rough lemon (C. jambhiri Lush) followed by that on Volkameriana (C. volkameriana Ten. & Pasq.), Yuma Ponderosa lemon (C. limon (L.) Burm. F.), sour orange (C. aurantium L.), Citrus macrophylla Wester, Morton citrange (C. sinensis cv. Washington navel × Poncirus trifoliata (L.) Raf.), Yuma citrange, Rangpur lime (C. limonia Obs.), Palestine sweet lime (C. limettioides Tan.), C-32 citrange and Citremon 1449 (C. limon × P. trifoliata), although no statistically significant differences were found between sour orange, the commercial rootstock used in Cyprus, and the other above mentioned rootstocks. Next in order as regards to total cumulative yield was Cleopatra mandarin (C. reticulata Blanko.) followed by Carrizo citrange, with statistically significant differences compared with sour orange. Trees on Citrumelo CPB-4475 (C. paradisi Macf. × P. trifoliata) and C-35 citrange died 3–4 years after grafting. Canopy volume was the lowest for trees on Cleopatra mandarin and Carrizo citrange. Yield efficiency A, expressed as total cumulative yield per trunk cross-sectional area, was the highest on C. macrophylla and lowest on Carrizo citrange. Rootstock significantly affected fruit size and weight, rind thickness, juice content, brix, total acids and brix:acid ratio. The results of the present study reveal that the most promising rootstocks that may replace sour orange for the local lemon variety ‘Lapithkiotiki’ under Cyprus conditions are Volkameriana, Yuma Ponderosa lemon, C. macrophylla and Citremon 1449.     

Towards crop improvement in bell pepper (Capsicum annuum L.): Transgenics (uid A::hpt II) by a tissue-culture-independent Agrobacterium-mediated in planta approach

Agrobacterium tumefaciens-mediated transformation has been one of the methods used to generate transgenic plants in bell pepper. An alternate transformation method that avoids/minimizes tissue culture would be beneficial for the improvement of bell pepper due to its recalcitrant nature. In this report, transgenic bell pepper plants have been developed by a tissue-culture-independent A. tumefaciens-mediated in planta transformation procedure. In the present study, two open pollinated varieties viz., Arka Gaurav and Arka Mohini were used for transformation. Agrobacterium strain EHA105 harboring the binary vector pCAMBIA1301 that carries the genes for β-glucuronidase (uid A) and hygromycin phosphotransferase II (hpt II) was used for transformation. GUS histochemical analysis of T₀ and T₁ plants at various stages of growth followed by molecular analysis using PCR, Southern analysis and RT-PCR allowed selection of transgenics. The method resulted in 17.8% and 11.4% of the T₀ plants in Arka Gaurav and Arka Mohini being selected as chimeric and 35.0% and 29.7%, respectively, were identified as stable transformants in the T₁ generation based on PCR analysis.     

A rapid,novel and high-throughput identification of putative bell pepper transformants generated through in planta transformation approach

The recent progress on bell pepper transformation through in planta approach is incredibly simple with high transformation efficiencies in indigenous varieties. This method produces chimeric primary transformants (T₀) hence, a large number of T₀ plants and an efficient high throughput screening strategy is necessary to be developed. In the present investigation, we have standardized a rapid and highly efficient method of screening putative bell pepper transformants. Bell pepper transformants harbouring gfp genes was developed using in planta approach and 1050 etiolated T₁ seedlings were screened using fluorescent microscope to select GFP positive plants. The molecular analysis such as, gene specific PCRs, semiquantitative RT-PCR and Southern blots analysis corroborated the transgene integration in selected T₁ transformants. Based on progressive selection strategy, the transformation efficiency was 27.4% in gfp screened plants. The fluorescence based screening is viable, rapid, requires minimal labour, expense, and expertise with less percent of non-transformant escapes; further, this method can be extended from bell pepper to other crops to identify putative transformants developed through in planta approach.     

Micro-grafting shoot-tip culture of citrus on three trifoliolate rootstocks

Simple techniques and growth regulator treatments to improve micro-grafting success of ‘Mexican’ lime, ‘Valencia’ orange and ‘Star Ruby’ grapefruit were studied. Most scion cultivars grafted best on ‘Carrizo’ citrange [Poncirus trifoliata (L.) Rat. x Citrus sinensis (L.) Osbeck]. Dipping the shoot-tip in 2,4-D or kinetin before grafting doubled the percentage of successful grafts. Highest grafting success occurred when the scion shoot-tip was placed in an inverted-T incision on the epicotyl of the rootstock.     

Genetic transformation of Prunus domestica L. using the hpt gene coding for hygromycin resistance as the selectable marker

The study and development of transformation technology with new selection schemes is important for various fundamental studies and for crop trait improvement via genetic engineering. Here we have shown that hygromycin resistance is an effective system for plum genetic transformation. Embryonic axes of mature seeds were co-cultivated with Agrobacterium strain LBA4404 containing the pC1381 plasmid carrying the hygromycin phosphotranferase gene (hpt) and β-glucuronidase (GUS) gene or with strain EHA105 containing the plasmid pC1301 carrying the same marker and reporter genes. The latter strain containing a pC2301 plasmid carrying the neomycin phosphotransferase gene (nptII) gene was used as a control. Infected explants were placed on shoot induction medium containing either 5 mg L⁻¹ hygromycin or 75 mg L⁻¹ kanamycin for selection. Green shoots developed from the explants under hygromycin pressure. These shoots showed continued and vigorous growth and development upon transfer onto fresh hygromycin medium. PCR using hpt sequence primers, and Southern blot analysis using a probe from the hpt gene, confirmed the presence of the transgenes and their stable integration in regenerated plants. Full transgenic plants were obtained in a greenhouse. Hygromycin selection was very effective and no escapes were observed. The study demonstrated that hygromycin resistance can be used as an effective selectable marker for plum transformation. The new system developed here is important and useful for multiple gene transformation in plum.     

Genetic transformation of Citrus sinensis cv. Hamlin with hrpN gene from Erwinia amylovora and evaluation of the transgenic lines for resistance to citrus canker

Transgenic Citrus sinensis (L.) Osb. cv. Hamlin plants expressing the hrpN gene were obtained by Agrobacterium tumefaciens (Smith and Towns) Conn-mediated transformation. hrpN encodes a harpin protein, which elicits the hypersensitive response and systemic acquired resistance in plants. The gene construct consisted of gst1, a pathogen-inducible promoter, a signal peptide for protein secretion to the apoplast, the selection genes nptII or aacC1 and the Nos terminator. The function of gst1 in citrus was evaluated in transgenic C. sinensis cv. Valencia harboring the reporter gene uidA (gus) driven by this promoter. Histochemical analysis for gus revealed that gst1 is activated in citrus leaves by both wounding and inoculation with Xanthomonas axonopodis Starr and Garces pv. citri (Hasse) Vauterin et al. Genetic transformation was confirmed by Southern blot hybridization in eight cv. Hamlin acclimatized plants. RT-PCR confirmed hrpN gene expression in seven cv. Hamlin transgenic lines before pathogen inoculation. Some hrpN transgenic lines showed severe leaf curling and abnormal growth. Six hrpN transgenic lines were propagated and evaluated for susceptibility to X. axonopodis pv. citri. RT-PCR confirmed gene expression in all six hrpN transgenic lines after pathogen inoculation. Several of the hrpN transgenic lines showed reduction in susceptibility to citrus canker as compared with non-transgenic plants. One hrpN transgenic line exhibited normal vegetative development and displayed very high resistance to the pathogen, estimated as up to 79% reduction in disease severity. This is the first report of genetic transformation of citrus using a pathogen-inducible promoter and the hrpN gene. Further evaluations of the transgenic plants under field conditions are planned. Nevertheless, the evidence to date suggests that the hrpN gene reduces the susceptibility of citrus plants to the canker disease.