Cloning,characterization and expression analysis of a 9-lipoxygenase gene in Gladiolus hybridus

Plant lipoxygenases (LOXs) is a functionally diverse class of dioxygenases involved in multiple physiological processes such as growth, senescence, and stress-related responses. Previously, based on the analysis of LOX1 activity and corm formation, it was depicted that LOX1 increased jasmonic acid (JA) synthesis and induced enlargement at top of stolon branches, forming cormels in Gladiolus hybridus. Here, LOX gene (GhLOX1) in full length was isolated from developing corms in G. hybridus. Sequence analysis showed that GhLOX1 was a novel LOX1 gene encoding 9-LOX. Transient expression analysis determined GhLOX1 to be a cytoplasm protein. Real time RT-PCR showed that the mRNA level of GhLOX1 gene correlated positively with LOX activity and was highest in cormels. GhLOX1 mRNA level, LOX activity and endogenous methyl jasmonate (MJ) content in corms increased steadily with a MJ gradient from 0.1 mmol/L to 0.5 mmol/L. Conversely, GhLOX1 mRNA level, LOX activity, and endogenous MJ content in corms steadily decreased with the increasing concentration of Salicylhydroxamic acid (SHAM). At the same time, we observed that MJ and SHAM treatments effected both LOX1 mRNA level and LOX activity, leading to promotion and inhibition of corm formation and enlargement, respectively. These data reveal that GhLOX1 is a novel LOX1 gene encoding a cytoplasm 9-LOX protein and its expression would contribute to LOX activity which plays an important role in corm formation and enlargement. In general, GhLOX1 is an ideal candidate for the promotion of corm formation and enlargement by gene manipulation in G. hybridus.     

水杨苷异羟肟酸和茉莉酸甲酯对唐菖蒲试管结球的影响

为探讨水杨苷异羟肟酸(SHAM)和茉莉酸甲酯(MJ)对唐菖蒲试管结球的影响,以'Rose Supreme'的籽球为外植体,研究了不同处理球茎内可溶性蛋白、内源MJ、碳水化合物(蔗糖、淀粉和纤维素)含量、脂氧合酶(LOX-1)、抗氧化酶(SOD、CAT、POD和APX)活性和LOX同工酶变化与试管结球之间的关系。结果表明：与对照相比,SHAM处理使结球时间明显推迟,结球率、球茎鲜样质量和直径降低;可溶性蛋白含量、LOX-1活性和MJ含量显著降低,LOX同工酶谱带变弱,并有2条同工酶带逐渐消失;抗氧化酶活性和碳水化合物含量降低。而MJ处理后上述指标均不同程度上调,0.5mmol·L^-1为测试浓度范围内促进唐菖蒲试管结球的最佳浓度。以上试验结果说明SHAM对唐菖蒲试管结球起抑制作用,而MJ有明显的促进作用,可能通过外施MJ提高了LOX活性及内源MJ含量,调动碳水化合物的积累,增加抗氧化酶的活性,从而对球茎的发生和生长进行调控。     

脂氧合酶对唐菖蒲籽球形成和膨大的影响

为探讨脂氧合酶(LOX-1)对唐菖蒲籽球形成和膨大的影响,以'Rose Supreme'和'Advanced Red'两个品种为试材,研究了籽球的生长发育与LOX-1活性,内源茉梨酸甲酯(MJ)、蔗糖、淀粉和纤维素含量变化之间的关系。结果表明,唐菖蒲叶片中测不到LOX-1活性,匍匐茎中LOX-1活性和内源MJ含量最高,籽球数量较多的品种'Rose Supreme'LOX-1活性和MJ含量显著高于'Advanced Red',说明LOX-1在唐菖蒲籽球形成和膨大过程中起着重要作用,其作用途径可能是通过影响茉莉酸类化合物(JAs)的生物合成对籽球的发生和生长进行调控。在唐菖蒲籽球生长发育过程中,新球和籽球中蔗糖、淀粉和纤维素含量都不同程度升高,并且与LOX-1活性呈正相关,说明蔗糖、淀粉和纤维素对唐菖蒲新球和籽球的形成和膨大起着重要作用。     

唐菖蒲脂氧合酶基因GhLOX1 对球茎膨大的影响

 以唐菖蒲（Gladiolus hybridus）品种‘Rose Supreme’的球茎为试材，应用Real time RT-PCR技术，分析唐菖蒲球茎在不同浓度的茉莉酸甲酯（methyl jasmonate，MJ）0.1、0.2、0.5 mmol · L^-1和水杨酸异羟肟酸（salicylhydroxamic acid，SHAM）0.5、0.75、1.0 mmol · L^-1处理后脂氧合酶（LOX）基因的的表达水平，并且对LOX活性、内源MJ含量以及球茎鲜样质量与体积进行测定。分析结果表明，GhLOX1表达水平、脂氧合酶活性、内源MJ含量以及球茎的鲜样质量与体积均随着MJ浓度的增加而逐渐提高。与此相反，经过不同浓度的LOX抑制剂SHAM处理后，以上各项指标随着SHAM浓度的增加而逐渐降低。同时，构建35S-GhLOX1过表达载体，利用根癌农杆菌介导法将GhLOX1导入马铃薯栽培品种‘中薯3号’的试管薯薄片中，转基因马铃薯植株块茎的数量、体积和鲜样质量分别得到了不同程度的提高。     

Identification of S-alleles in pear (Pyrus communis L.) cv. ‘Rocha’ and other European cultivars

In this work we report the cloning and identification of S-RNase alleles responsible for gametophytic self-incompatibility (GSI) of ‘Rocha’ pear and of 13 other European pear cultivars that might be used as its pollinators. Partial sequences of S-RNase alleles were amplified by PCR with specific primers hybridising in conserved regions of previously identified S-RNase alleles of Pyrus communis, cloned and sequenced and the S-genotype of eight pear cultivars was fully determined. Three cultivars (‘General Léclerc’ (SqSl), ‘Tosca’ (SbSl) and ‘Alexandrine Douillard’ (SbSk)) shared no S-alleles with ‘Rocha’ (SaSe) and shall be totally compatible with this cultivar. None of the cultivars analysed showed an identical amplification pattern to the one observed in ‘Rocha’, so the other cultivars shall be at least semi-compatible. One new allele was identified in P. communis cv. ‘Beurré d’Avril’ (designated as St). The determination of both S-RNase alleles of cvs ‘Rocha’, ‘Beurré Precoce Morettini’ (SeSk) and ‘Tosca’ and the identification of one S-RNase allele in cvs ‘Carapinheira’ (Sb), ‘Amêndoa’ (Se), ‘Pérola’ (Sk) and ‘Beurré d’Avril’ (St) are important contributions for the effort recently developed worldwide to establish groups of sexual compatibility among European pears.     

Responses of eight chile peppers to saline water irrigation

Salt tolerance of five cultivars of Capsicum annuum L. Early Jalapeno, Golden Treasure, NuMex Sweet, NuMex Joe E. Parker, and Santa Fe Grande, two cultivars of C. chinense Jacq. Habanero and Pimienta De Chiera, and one accession of C. annuum, NMCA 10652, were evaluated in a field study. Seedlings were transplanted in late May to field raised beds containing loamy sand soils in a semi-arid environment. Plants were well irrigated throughout the experiment. Three saline solution treatments, prepared by adding NaCl, MgSO₄, and CaCl₂ to tap water at different amounts to create three salinity levels of 0.82 dS m⁻¹ (control, tap water), 2.5 dS m⁻¹, and 4.1 dS m⁻¹ electrical conductivity (EC), were initiated on 15th June and ended in late August. Among the eight varieties, NMCA 10652 had the highest survival percentage at 100% in the 4.1 dS m⁻¹ treatment, followed by ‘Early Jalapeno’, ‘NuMex Sweet’, ‘Pimienta De Chiera’, ‘Santa Fe Grande’, ‘Golden Treasure’, and ‘NuMex Joe E. Parker’. ‘Habanero’ had the lowest survival at 28%. Compared to control, final shoot dry weight of the plants irrigated with saline solution at 4.1 dS m⁻¹ was reduced by 92% in ‘Habanero’, followed by ‘Golden Treasure’ at 80%. For fruit fresh weight in 4.1 dS m⁻¹ vs. control, ‘Habanero’ had the highest reduction at 86%, followed by ‘Golden Treasure’ at 74%, while NMCA 10652 and ‘Santa Fe Grande’ had the least at 26% and 19%, respectively. NMCA 10652, the most tolerant to salinity, had the lowest leaf Na⁺ accumulation, while ‘Habanero’, the most sensitive to salinity, had the highest Na⁺ in the leaves. For leaf Cl⁻, ‘Early Jalapeno’ had the highest, while ‘Habanero’ had the lowest Cl⁻ accumulation in the leaves. Generally, sensitive varieties accumulated more Na⁺ and/or Cl⁻ in leaves, except for ‘Early Jalapeno’, which was relatively tolerant to salinity but had high Na⁺ and Cl⁻ accumulation in leaves.     

Differential structure and physiological response to canker challenge between ‘Meiwa’ kumquat and ‘Newhall’ navel orange with contrasting resistance

Asiatic citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most detrimental diseases affecting citrus production. ‘Meiwa’ kumquat (Fortunella crassifolia) and ‘Newhall’ navel orange (Citrus sinensis) are two genotypes with contrasting Xcc resistance. In the current study, efforts were made to perform pairwise comparison between them in terms of structural difference and biochemical response to Xcc attack. Pinprick inoculation demonstrated that ‘Meiwa’ had lower disease incidence rate and smaller lesion size, concurrent with significantly less bacterial number at the inoculation sites relative to ‘Newhall’. Microscopic observation demonstrated that stomatal density and size of ‘Meiwa’ were significantly smaller than ‘Newhall’. Stomatal apertures of ‘Meiwa’ were smaller than ‘Newhall’ measured at 2 h after their leaves were immersed in Xcc suspension, consistent with the bacteria number in the internal tissues counted at 4 d. Epicuticular wax content of ‘Meiwa’ was significantly higher than that of ‘Newhall’, while no difference in palisade and spongy tissue was observed between them. Enzymatic activity assay showed that after Xcc inoculation, ‘Meiwa’ exhibited significantly higher activities of catalase (CAT), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) in comparison with ‘Newhall’ at most of the sampling time points. Our data showed that the two genotypes with contrasting Xcc resistance exhibited clear difference in the structure and physiological response, which may provide information to facilitate our understanding of Xcc resistance.     

Effect of synthetic auxins on fruit size of five cultivars of Japanese plum (Prunus salicina Lindl.)

Most of the Japanese plum (Prunus salicina) cultivars grown in Israel produce relatively small fruit. Application of 2 l solution tree⁻¹ of 25 mg l⁻¹ 2,4-dichlorophenoxypropionic acid (2,4-DP) as butoxyethyl ester (Power™), 15 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid (3,5,6-TPA) as free acid (Maxim^®), or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) + 30 mg l⁻¹ naphthaleneacetic acid (NAA) (0.3% Amigo™) at the beginning of pit-hardening, when fruitlet diameter was ca. 22 mm, caused an appreciable and significant increase in fruit size. The yield of large fruit per cv.: ‘Kesselmen’ (100% increase), ‘Songold’ (100%), ‘Black Diamond’ (800%), ‘Royal Diamond’ (160%) and ‘Royal Zee’ (100%). As a result, the total yield of all five cultivars was also increased dramatically. Anatomical studies with ‘Songold’ revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect either on fruit quality at harvest (and after 1 week in shelf-life), or on return yield in the following year.     

Cybrid/hybrid plants regenerated from somatic fusions between male sterile Satsuma mandarin and seedy tangelos

Somatic hybridization provides an alternative for transferring mitochondria-encoded cytoplasmic male sterility (CMS). Herein, symmetric protoplast electrofusion was conducted between embryogenic callus protoplasts of Citrus unshiu Marc. cv. Guoqing No. 1 (G1), a CMS cultivar, and mesophyll-derived protoplasts of seedy ‘Page’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)] or ‘Nova’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)], to transfer CMS trait. Flow cytometry analysis showed that 14 plants recovered from G1 + ‘Page’ tangelo that displayed typical morphological character of ‘Page’ were diploid, and 6 plants regenerated from G1 + ‘Nova’ tangelo were tetraploid. Genetic compositions of regenerated plants from the two fusions were determined by SSR, CAPS and chloroplast-SSR analysis. Cybrid nature of diploids from G1 + ‘Page’ tangelo with nuclear DNA from ‘Page’, mitochondrial DNA (mtDNA) from the G1 and chloroplast DNA (cpDNA) derived from either parent was confirmed. Tetraploid plants from G1 + ‘Nova’ tangelo were identified as somatic hybrids with random cpDNA inheritance. The regenerated cybrid and hybrid plants hold great potential for Citrus seedless breeding at diploid or triploid levels.     

Cellular wall metabolism in citrus fruit pericarp and its relation to creasing fruit rate

Citrus creasing results in serious economic loss in many citrus orchards. Based on the different incidence of creasing, two sweet orange cultivars ‘Hong Jiang’ (grafting chimaera of which flesh is mostly from Citrus reticulata Blanco and peel is from Citrus sinensis Osbeck in) and ‘An Liu’ (Citrus sinensis Osbeck) were used to investigate the creasing rate. The ultrastructure in cellular wall, cellular wall component, cellular wall degradation-related enzymes and expansin (Ct-Exp1) of sweet orange during fruit ripening in field were comparatively analyzed. ‘Hong Jiang’ sweet orange had a higher creasing rate than ‘An Liu’ during ripening. The activities of polygalacturonase, cellulase and pectinesterase in cellular wall of ‘Hong Jiang’ increased more markedly compared with ‘An Liu’. The increases in the content of soluble pectin, ionically associated pectin, covalently bound pectin, hemicellulose and cellulose of ‘Hong Jiang’ were higher than those of ‘An Liu’. Furthermore, the enhanced degradation of the ultrastructure in the albedo cellular walls was observed in ‘Hong Jiang’. Moreover, the northern blot analysis indicated that the Ct-Exp1 gene expressed more strongly in peel of ‘Hong Jiang’ than ‘An Liu’ during fruit ripening. These data suggest that enhanced loss of pectin and cellulose in the cellular walls of peel tissue of sweet orange could result in fruit creasing.     

6-Benzylamino purine stimulates in vitro shoot organogenesis in Eucalyptus erythronema, E. stricklandii and their interspecific hybrids

In vitro bud and shoot organogenesis was investigated for the ornamental plants Eucalyptus erythronema var. erythronema, E. stricklandii and their interspecific hybrids cv. ‘Urrbrae Gem’ and ‘Hybrid 2.5’ by using 0.0, 0.1, 0.25, 0.5 or 1.0 μM BAP on apex and leaf explants. Callus developed on all explants and increased with all concentrations of BAP without significant differences between BAP concentrations. Buds formed on apex and leaf explants of E. erythronema and E. cv. ‘Urrbrae Gem’ especially with 1.0 μM BAP, but these buds rarely developed into shoots. Bud clusters formed on E. erythronema and E. cv. ‘Urrbrae Gem’ apex and leaf explants whereas E. stricklandii and ‘Hybrid 2.5’ produced fewer, individual buds on the explant. Shoots regenerated from apex explants of all genotypes with all levels of BAP, whereas few shoots of any genotype regenerated from leaf explants regardless of the number of buds formed. Shoots from apex explants could be multiplied successfully. Light microscopy showed meristems developed within the callus, and at the callus and bud surfaces. However, few shoots developed considering the level of bud and meristem formation. This report is the first for successful shoot organogenesis and multiplication in an ornamental eucalypt.     

Syrian pear (Pyrus syriaca) as a pollinator for European pear (Pyrus communis) cultivars

In Israel four European pear cultivars are grown: ‘Spadona’ is the main cultivar and ‘Coscia’, ‘Gentile’ and ‘Spadochina’ are its pollinators. However, molecular S-genotyping revealed that ‘Spadona’ is semi-compatible with its three pollinators. This explains, at least in part, the relatively low pear yield in Israel. The Syrian pear (Pyrus syriaca) grows wild in Israel and blooms intensively, overlapping the blooming of the cultivated European pears. Cross-fertilization between Syrian pear and ‘Spadona’ was shown to be efficient suggesting that Syrian pear might be a potent pollinator for ‘Spadona’. Twenty-six Syrian pear seedlings, from different sites in north-east Israel were S-genotyped identifying 11 that are fully compatible with the four European pear varieties cultivated in Israel. By this screening, 24 different S-RNases were cloned; ten of them are new, whereas the other fourteen had been identified previously. In addition, seedlings of two wild pear species were also S-genotyped. Two seedlings from Pyrus betulifolia and one from Pyrus korshinskii were found to be genetically compatible with the four European pear cultivars. From these seedlings four S-RNases were cloned, two are new, one had been cloned previously and one was identical to an S-RNase allele cloned from Syrian pear in this work.     

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.     

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.     

Effect of the expression level of an AGAMOUS-like gene on the petaloidy of stamens in the double-flowered lily, ‘Elodie’

Double-flowered lilies, in which stamens are converted to petaloid organs, are valuable for horticulturists. ‘Elodie’ is a double-flowered lily cultivar in which stamens are homeotically converted into petaloid organs in whorl 3. The ‘Elodie’ cultivar shows individual variation in stamen structure and it was therefore classified into the following three types according to the strength of petaloidy of the stamens: weak (type-I), intermediate (type-II), and strong (type-III) phenotypes. The AGAMOUS (AG) gene is a class C floral identity gene in Arabidopsis thaliana that is involved in the formation of stamens and carpels. An AG-like gene was isolated from ‘Elodie’ (LelAG1) and its expression was compared between flower types. The LelAG1 gene was expressed in whorls 3 and 4, but not in whorls 1 and 2 in all flower types. In type-I flowers, LelAG1 was expressed strongly in whorls 3 and 4, while its expression was significantly decreased in whorl 3 in type-III flowers. In type-II intermediate phenotype flowers, the expression level of LelAG1 in whorl 3 was reduced by 60%. These results suggest that the expression level of AG-like genes is correlated with the degree of petaloidy of the stamens.     

The physiological responses of carnation cut flowers to exogenous nitric oxide

Nitric oxide (NO) is a highly reactive signaling molecule and plays a variety of physiological roles in plants. The research on the application of NO to postharvest preservation of flowers and fruits shows great promise in recent years. However, the physiological mechanism of exogenous NO to affect cut flowers is not very clear. Therefore, an experiment was conducted to study the effect of exogenous NO on the vase life and physiological basis of Dianthus caryophyllus L. variety ‘Monte’. In the present study, 0.1 mmol L⁻¹ sodium nitroprussiate (SNP) was used as the NO donor, and 5 μmol L⁻¹ methylene blue (MB-1) was used as its scavenger based on the preliminary experiment. We evaluated the physiological index including increase of stem weight, malondialdehyde (MDA) concentration and lipoxygenase (LOX) activity, and cell protection enzymes activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX). The results showed that exogenous NO could significantly extend the vase life of cut carnation flowers and markedly increase fresh mass. The balance of water metabolism and the activities of SOD, POD, CAT and APX also showed improvement, while the production of MDA content and LOX activity were obviously decreased. The results suggest that exogenous NO could delay petal wilting in carination cut flowers, maintain water metabolism, the antioxidative enzymes activity and mass-eliminate reactive oxygen species (ROS) and as well as cell membrane stability. Moreover, the results indicated that MB-1 had the ability to reverse the active effects of NO on different physiological indexes. Therefore, the vase life of cut carnation flowers was markedly extended by SNP treatment.     

Acid invertase as a serious candidate to control the balance sucrose versus (glucose + fructose) of banana fruit during ripening

Six banana varieties: 3 ‘dessert’ ones: ‘IDN 110’; ‘Kirun’; and ‘Grande Naine’, and 3 ‘cooking’ ones: ‘Galéo’; ‘Sowmuk’; and ‘French’ were used to investigate the relationship between sugar profiles and activities of sucrose-phosphate synthase (SPS, EC 2.4.1.14), sucrose synthase (SuSy, EC 2.4.1.13) working in the hydrolytic way, invertases (EC 3.2.1.26) (neutral (NIV) and acid (AIV)). Expression of a Musa cell-wall invertase (MaCwINV1/pBANUU103) gene was additionally studied during fruit development and ripening ex planta after ethylene treatment of two of these varieties. Close amounts of soluble sugars (sucrose + glucose + fructose) were measured in the different varieties at ripe stage. SPS activity was either almost constant or increased continuously or transitorily during ripening of all varieties, concomitantly to total soluble sugar (sucrose + glucose + fructose) accumulation. Together with previous data obtained from ‘Cavendish’, our data lead us, (i) to strengthen the hypothesis that this enzyme is likely to have a major role in the synthesis of sucrose during ripening of banana and (ii) to underline the complexity of the mode of SPS activity regulation already pointed out. Interestingly, for the first time in banana, two diploid and cooking varieties: ‘Galéo’ and ‘Sowmuk’ were found almost sucrose-free, in good agreement with a 6.4-fold higher mostly vacuolar AIV activity than that of the two desserts and diploid ones. Otherwise, expression of a MaCwINV1/pBANUU103 (cell wall) gene was followed in the two most contrasted varieties in matter of sucrose content: ‘Sowmuk’ almost sucrose-free, and ‘IDN 110’ accumulating high level of sucrose. Compared to ‘IDN110’, the mRNA level of MaCwINV1/pBANUU103 gene was higher (up to 173-fold) in ‘Sowmuk’ concomitantly with the low level of sucrose of ‘Sowmuk’. Our data let us to conclude that the AIV is probably one of the main determinants involved in the regulation of sucrose level during banana fruit ripening, even if the form, vacuolar or cell wall-linked is not determined yet. Otherwise, the MaCwINV1/pBANUU103 gene appears as a putative candidate gene that could contribute to regulate this level.     

Pollen biology of ornamental ginger (Hedychium spp. J. Koenig)

An improved in vitro pollen germination assay was developed to assess the viability of stored Hedychium pollen. The effect of polyethylene glycol (PEG) (10, 15, and 20%, w/v) on pollen germination and tube growth was evaluated for Hedychium longicornutum and two commercial Hedychium cultivars, ‘Orange Brush’ and ‘Filigree’. Overall, the inclusion of PEG 4000 in the medium improved both pollen germination and tube growth for the three different genotypes tested and the results varied depending on genotype. In vitro germination was used to assess the viability of Hedychium pollen stored up to two months. Pollen nucleus status was determined for four Hedychium cultivars, ‘Orange Brush’, ‘Anne Bishop’, ‘Filigree’, and ‘Daniel Weeks’. Pollens of ‘Orange Brush’, ‘Anne Bishop’, and ‘Daniel Weeks’ were found to be binucleate but ‘Filigree’ was shown to possess both binucleate and trinucleate pollens. High pollen:ovule ratio values were obtained in several Hedychium taxa. The results obtained on the nuclear pollen status and pollen:ovule ratios will further our understanding of the pollination biology and help clarify the taxonomy and phylogeny of Hedychium species.     

Anthocyanins from flowers of Hippeastrum cultivars

The anthocyanins, cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (1) and pelargonidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (2), were isolated from the ornamental flowers of a Ugandan Hippeastrum cultivar by a combination of chromatographic techniques, and their structures were elucidated mainly by the use of homo- and heteronuclear nuclear magnetic resonance spectroscopy and electrospray mass spectrometry. The same anthocyanins were found in six different Hippeastrum cultivars purchased in Norway. However, the absolute amount of the anthocyanins (0.08–1.79 mg/g, fresh weight) and the relative proportions of the individual anthocyanins varied from cultivar to cultivar (13.2–96.5% of 1). The colours of the fresh petals of the three cultivars ‘Red Lion’, ‘Royal Velvet’ and ‘Liberty’ were described by the CIELab coordinates L^* (lightness), C^* (chroma) and h_ab (hue angles). All the cultivars showed hue angles corresponding to scarlet nuances (h_ab = 22–35°), with the highest value in ‘Red Lion’. The most reddish petals (in ‘Royal Velvet’) contained the highest relative proportion of 1. Thus, the in vivo colours of these cultivars seem to be correlated with the relative proportions of individual anthocyanin in the petals.