Effects of lipoxygenase on the corm formation and enlargement in Gladiolus hybridus

To investigate the effects of lipoxygenase (LOX1) on the corm formation and enlargement in Gladiolus hybridus, the field and in vitro researches were conducted to detect the growth and development course, the LOX1 activity, the contents of endogenetic methyl jasmonate (MJ), sucrose, starch and cellulose, as well as LOX isozymes and the expression of its genes in ‘Rose Supreme’ and ‘Advanced Red’. The field investigation showed that the highest activities of LOX1 activity and MJ content in stolons, but not detected in leaves, which were higher in ‘RS’ than in ‘AR’. And the contents of sucrose, starch and cellulose in corm and cormels showed different increase during growth and development of Gladiolus corm. The in vitro investigation showed that the salicylhydroxamic acid (SHAM) treatment caused obviously delayed corm formation, decrease of fresh weight and diameter, and percentage of corm formation; reduced contents of total dissoluble protein, MJ and carbohydrates, and LOX1 activity. Additionally the activity of LOX isozymes became weaker, with two isozyme bands disappeared. In contrast, these physiological and biochemical indexes were increased under MJ treatment. RT-PCR also demonstrated that the expression of LOX1 gene was inhibited under SHAM treatment, induced under MJ treatment. These results suggested that LOX1 might regulate the growth and development of Gladiolus corm through affecting JAs biosynthesis cycles, and then resulting in carbohydrate accumulation. The corm formation could be improved by MJ, but suppressed by SHAM, in which, MJ 0.5 mmol/L was the optimum concentration for promoting corm formation and enlargement in Gladiolus.     

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。     

唐菖蒲茉莉酸生物合成关键酶基因LOX1的克隆及表达分析

 以唐菖蒲（Gladiolus hybridus）品种‘Rose Supreme’的球茎为试材,通过RT-PCR和RACE技术克隆到了一个全长为2 797 bp的茉莉酸（Jasmonic acid,JA）生物合成关键酶LOX基因的cDNA序列,命名为GhLOX1,属于9-LOX。该序列含有一个2 541 bp的开放阅读框（ORF）,编码846个氨基酸,推导的蛋白质分子量为94.90 kD。RT-PCR表达分析表明,该基因在唐菖蒲叶、花、根、匍匐茎、新球茎和籽球上都表达,而在叶和匍匐茎中表达量最高,在花和籽球中表达量较低;离体条件下,经过0、0.1、0.5、1.0、2.0 mmol · L^-1的不同浓度梯度的水杨酸（SA）处理后,其表达水平随着浓度的升高而降低,当SA浓度达到2.0 mmol · L-1时没有表达。     

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。     

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。     

异源表达唐菖蒲GhOPR3 提高了拟南芥的抗逆性

 通过RT-PCR 与RACE 技术从唐菖蒲（Gladiolus hybridus）品种‘Rose Supreme’球茎中克 隆茉莉酸生物合成途径中的关键酶——12–氧–植物二烯酸还原酶（12-oxo-phytodienoic acid reductase， OPR）基因的1 489 bp 全长cDNA 序列，quantitative RT-PCR 结果表明，GhOPR3 在唐菖蒲叶、花、根、 匍匐茎、新球茎和籽球中都表达，其中在籽球和匍匐茎中相对表达量较高；0.1 ~ 0.5 mmol · L-1 的茉莉酸 甲酯（Methyl jasmonate，MJ）处理后，提高了GhOPR3 在球茎中的表达量和内源MJ 含量；采用农杆菌 介导侵染拟南芥花粉，进行GhOPR3 基因的过表达分析，过表达拟南芥株系较野生型提高了耐盐性和抗 旱性；提高了机械损伤后相关基因的表达水平和内源MJ 含量。     

Effect of 2-Chloroethanephosphonic Acid (Ethrel) on Health,Dormancy, and Flower and Corm Yield of Gladioli

2-Chloroethanephosphonic acid (Ethrel), an ethylene releasing compound, was used on gladiolus corms as a 30-minute dip treatment in various concentrations and at various times during storage.

The most important effect of Ethrel was the marked increase in health and survival of treated plants infected with latent Fusarium.

Ethrel at 1000 ppm enhanced sprouting when corms were stored at high temperatures or stored for a short period at 5 °C. Treatment of cold-stored corms at high concentrations delayed sprouting.

Ethrel increased corm splitting, delayed flowering and slightly shortened flower stems. The size of harvested corms was reduced, but the yield of cormels was greatly increased by Ethrel.     

The mode of action of ethephon in increasing health of gladiolus corms

Gladiolus corms and cormels were treated with various combinations of ethephon and the fungicides methylethyl mercury benzoate, benomyl, thiobendazole and methylmercury dicyandiamide either before or after storage at 10 °C and 30 °C for 6 weeks. The treated corms were examined for germination, splitting and rot symptoms caused by Fusarium oxysporum f. sp. gladioli.

Ethephon increased corm splitting in all treatments; it also increased germination in corms stored at 30 °C, and it increased the proportion of corms showing typical Fusarium rot symptoms by stimulating the growth of latent internal Fusarium. The effectiveness of the fungicides tested against F. oxysporum f. sp. gladioli was enhanced by the addition of ethephon. Permeability of corms to methylethyl mercury benzoate was increased by ethephon, and it is concluded that ethephon improves the efficiency of this fungicide, and probably of other fungicides as well, by increasing their penetration into gladiolus corms.     

Prediction of saffron crocus corm mass by geometrical attributes

Saffron (Crocus sativus L.) is the most precious spice plant in the world. The only method for the reproduction of saffron is cultivation of corm. It is necessary to know relationships between mass and size of saffron corms in order to design and develop of planting, harvesting, grading and handling equipments. The objective of this research was to determine the most accurate models to predict saffron corm mass based on geometrical attributes. Models for predicting the mass of corms were classified as follows: (1) single or multiple variable regressions of saffron corm mass and dimensional characteristics, (2) single or multiple variable regressions of saffron corm mass and projected areas and (3) single regression of saffron corm mass based on its actual volume and calculated volume assuming the saffron corm shapes of sphere, oblate spheroid and ellipsoid. Three sets of corms were considered from Torbat-e-Heydareyeh, Kashmar, and Gonabad fields in Khorasan province of Iran. Some physical characteristics that were needed for the saffron corm mass modelling were determined for the three regions corms. The results showed that there were not significant differences between mean values of mass, height, major diameter, geometric mean diameter and projected areas of saffron corms for the three regions. However, there were significant differences between saffron corm minor diameter, volumes, corm density, bulk density and porosity for the three regions. Therefore, the mass of the corms was modelled based on the height, major diameter and projected areas irrespective to the regions, whereas in the third classification models, the mass of the corms was modelled for each region. The statistically significant fitted models were selected based on the higher coefficient of determination (r²) and lower regression standard error (RSE). The results of mass modelling showed that the prediction of saffron corm mass based on the major diameter (M = 0.0017a^2.5168, r² = 0.94, p < 0.05) and the first projected area (M = 0.0019 (PA₁)^1.2916, r² = 0.96, p < 0.05) were the most appropriate models. The model that predicted the mass of saffron corm based on the estimated volume of an ellipsoid shape was found to be the most appropriate. The developed models and data of physical characteristics of saffron corms could be used for designing and developing the saffron corm grader and other cultivation equipments.     

In vitro corm production in Gloriosa superba L., an Ayurvedic medicinal plant

Summary

In vitro production of corms from Gloriosa superba L. using three kinds of explants: dormant, non-dormant corm buds and 30 d old in vitro derived multiple shoots is reported. Excellent responses was obtained in terms of corm production using MS medium supplemented with B₅ vitamins, 6% sucrose, 2iP, ADS and ANL for corm formation from dormant corm buds, using Kin, ADS and ANL for corm formation from non-dormant corm buds and using BAP, ADS and ANL for callus derived multiple shoot corm formation. Well developed corms were obtained from the tissue cultured plants with no dormancy breaking requirement.     

Corm induction and multiplication of Amorphophallus albus in vitro

Summary

Amorphophallus albus, belonging to the family Araceae, has attracted widespread attention due to its considerable economic and medicinal importance. The natural propagation coefficient of A. albus is very low, which limits application of this crop. In vitro corms can be used for propagation of A. albus and have been proved to be superior over in vitro plantlets. To optimise procedures for in vitro corm production and multiplication, the effects of phytohormones, sucrose concentrations and incubation conditions with desirable phytohormone combinations for callus induction, corm formation and corm growth of A. albus were investigated. The results showed that calli were induced at high frequency from petiole segments on Murashige and Skoog medium supplemented with 1.0 mg l^–1 naphthaleneacetic acid (NAA) and 1.0 mg l^–1 6-benzyladenine (BA). Compact nodular calli were desirable for corm formation, and optimum corm formation was obtained in the presence of 0.5 mg l^–1 NAA and 2.0 mg l^–1 BA. With this auxin and cytokinin combination, an increase in sucrose concentration from 2% to 6% (w/v) significantly increased the corm formation rate and favoured corm growth, but negative effects occurred at higher sucrose concentrations. By incubating over a range of temperatures from 19°C – 28°C, 22°C produced the largest numbers of corms and highest mean fresh weight of each corm. Short-day (8 h) or long-day (16 h) photoperiods did not affect corm formation and growth significantly, except that corm weight fell under long-day conditions. The multiplication rate of in vitro corms was enhanced by apical meristem wounding. It was possible to store in vitro produced corms at 4°C for as long as 90 d to overcome apical dormancy and accelerate sprouting after planting into soil. This work has established an efficient protocol for multiplication of A. albus through an in vitro corm system.     

Chemical composition of nine edible aroid cultivars of Bangladesh

Leaf blades, petioles, corms and cormels of 9 cultivars of edible aroids were analyzed to determine the content of protein, carbohydrate, crude fibre, ether extract, ash, carotene, thiamine, riboflavin, ascorbic acid, iron, calcium and phosphorus. The cultivars represent the genera Colocasia, Xanthosoma, Alocasia and Amorphophallus. Wide differences among the cultivars in respect of almost all of the components of analysis were observed.     

The influence of illumination levels of daylength extension on yield of winter-grown gladioli in Queensland

The yields of inflorescences, corms and cormlets, and inflorescence quality of winter gladioli were studied under varying illumination levels of 12 hour daylength extension ranging from 0 to 333 lux. The illumination level of the daylength extension for each plot depended on its distance from 2 tungsten halogen light sources mounted at 6 m above ground on poles spaced 8 m apart. The lamps were adjusted for wide beam distribution and aimed at points on the ground 8 m from the base of the poles.Inflorescence yield was increased by increasing the illumination level of daylength extension. The relationship between horizontal illumination level in lux (x) and number of inflorescences (y) was described byy=27.3?10.9 exp(?0.02x)Ninety-seven % of maximum inflorescence yield was obtained at a daylength extension illuminance of 144 lux, while 97% of the maximum number of florets per spike and the other flower quality characteristics were obtained at a daylength extension illumination level of approximately 100 lux. Ninety-seven % of the maximum number of days to flowering was approached at a daylength extension illuminance of 45 lux.There was no clear relationship between the illumination level of daylength extension and number and weight of new corms, or the average weight of each new corm. However, the weights of cormlets per plot and per new corm were higher from plants which received no daylength extension.The commercial application of tungsten halogen lamps to provide daylength extension for winter gladioli production is discussed.     

芋淀粉合成酶AGPase基因的克隆及表达分析

通过对‘靖江香沙芋’转录组数据进行功能注释,克隆获得‘靖江香沙芋’淀粉合成酶AGPase的基因,序列全长为1 596 bp,命名为CeApS1（NCBI：Colocasia KU288757,未公布）。该基因编码532个氨基酸,蛋白含有1个NTP_transf_3结构域（Gly¹⁰⁴ ~ Pro³⁰⁶）和1个PbH1结构域（Gly⁴⁷³ ~ Ser⁵¹⁵）,编码产生AGPase小亚基,等电点和分子量分别为6.73和57.6 kD。ApS1在单子叶和双子叶植物中广泛存在,CeApS1与紫萍（AIZ00992.1）中相应蛋白的亲缘关系最近,相似性为83.4%,与草莓（AAS00541.1）、苜蓿（XP_003617925.1）、水稻（AAK27313.1）和小麦（CAA46879.1）等植物中相应蛋白的亲缘关系较远。CeApS1在‘靖江香沙芋’的母芋和子孙芋中表达量较高,在叶片、叶柄和根中低表达。芋球茎中的CeApS1表达水平与球茎发育密切相关,在播种160 d后达到最大值。该基因的表达模式与球茎淀粉含量的积累显著正相关。     

Effect of low temperature on breaking dormancy and flowering of Arisaema sikokianum (Araceae)

Arisaema sikokianum (Araceae) native to Japan is classified as a vulnerable species in the Red Data Book of Japan. Control of dormancy is essential for efficient corm production and forcing culture. Sprouting of both vegetative and reproductive corms was enhanced by exposure to low temperature. Vegetative corms exposed to low temperatures at 5 °C longer sprouted faster when grown at 20 °C. Effective temperatures for breaking dormancy was 5 °C. Reproductive corms treated at 5 °C longer showed shorter days to flower. Successful forcing culture was achieved; corms treated at 5 °C from November for 30 days flowered on 5th February.     

Response of gladiolus (Gladiolus spp) plants after exposure corms to chitosan and hot water treatments

The state of Morelos, Mexico has gradually become an important producer of gladiolus. Some preconditioning treatments of corms are empirically done causing uneven emergence and low quality of flowers. In this investigation, before planting, gladiolus corms var. ‘Blanca Borrego’ were dipped in chitosan (chitosan reagent and commercial chitosan Biorend^®), in hot water at various temperatures and in treatments combined with Biorend^® at 1.5% and hot water. Results indicated that the most influenced variables were corm germination, number of flowers per spike, number of cormlets and vase life. Overall, the commercial product Biorend^® at 1.5% accelerated corm emergence in approximately 4 days, the number of flowers increased by 2–7 and the vase life extended for 3 days. The number of cormlets was also duplicated. Corms dipped in the commercial chitosan Biorend^® at 1.5% at different intervals of time were not greatly affected except for the emergence and number of cormlets. However, for this experiment there were significant effects on the number of leaves and flowers because of the interactions between chitosan and the immersion time. The temperature of 55 °C affected plant development because emergence was delayed by 6 days; and there were less number of leaves, flowers and cormlets. On the other hand, the incidence of Fusarium oxysporum in naturally infected corms was 0% at temperatures of 55 °C and 50 °C. Immersion times (0, 10, 15 and 20 min) in hot water at 50 °C did not show significant effects on plant development and vase life. Corms dipped in Biorend^® at 1.5% and hot water at 50 °C accelerated their emergence for about 1–7 days, the number of flowers increased by two, extended the storage life for 1–3 days and increased the number of cormlets. The integration of these two treatments -Biorend^® and hot water- might be a good option for increasing the gladiolus plant quality and vase life.     

Fruit calcium content and lipoxygenase activity in relation to albinism disorder in strawberry

Nutrient elements and lipoxygenase (LOX) activity were determined in strawberry fruit to establish a relationship, if it exists, between nutrient ratios (N/Ca and K/Ca), and lipoxygenase activity with albinism disorder. About 33% strawberry fruit were affected by albinism. Etna had highest incidence of albinism (48.6%) and Sweet Charlie the lowest (16.2%). Dry matter content (%) was lower in albino fruit (5.23%) than normal fruit (7.36%). The concentration of N, P, and Mg did not differ significantly, but that of K (1.87 mg g⁻¹ fresh weight) was notably higher and of Ca (0.105 mg g⁻¹ fresh weight) was lower in albino fruit than normal fruit. Consequently, the nutrient ratios, N/Ca (9.78) and K/Ca (16.96) were higher in albino fruit than normal fruit. Cultivars differed widely in respect to dry matter (%), mineral content and nutrient ratios. LOX activity determined on dry weight or fresh weight basis was significantly higher in albino fruit than normal fruit, with significant differences among cultivars. Positive correlations existed between nutrient ratios and albinism incidence (r = +0.338), LOX activity and albinism incidence (r = +0.412), and LOX and nutrient ratios (r = +0.448). Thus, it appears from the study that calcium and LOX activity may not the basic cause of albinism in strawberry, but these may be involved in senescence or fruit ripening process, as LOX activity was lower in albino than in normal fruit.     

芋种质资源染色体倍性鉴定

利用流式细胞术（Flow cytometry,FCM）,对芋属（Colocasia）中种质资源类型最为丰富的滇南芋（C.?antiquorum Schott）和芋〔C.?esculenta（L.）Schott〕的染色体倍性进行了鉴定。结果表明：滇南芋的染色体倍性为2n=2x=28。芋中魁芋的染色体倍性表现为2n=2x=28|多头芋、魁子兼用芋为2n=3x=42|多子芋一般为2n=3x=42,但白芽乌绿柄多子芋为2n=2x=28,另外来自印度的绿柄多子芋也为2n=2x=28。     

不同类型魔芋种芋的生长分析

通过对不同类型魔芋脱毒种芋的萌芽出苗特性、田间长势、分苗数、田间发病情况、产量、球茎膨大倍数、繁殖系数、生长周期等进行研究。研究结果表明,一级包衣种、二级种、三级种比原原种提前8～18天出苗;一级包衣种、二级种的分苗数较多;一级包衣种、二级种、三级种的田间发病率低于原原种,其中一级包衣种发病率最低,仅为7.4％。二级种、三级种综合性状较好,生产力水平高,膨大倍数高,可通过规模化生产得到大量种芋,解决生产中种芋缺乏的问题。