Anthocyanin accumulation and changes in activities of phenylalanine ammonia-lyase and chalcone synthase in roselle (Hibiscus sabdariffa L.) callus cultures

Time-course changes in anthocyanin accumulation, phenylalanine ammonia-lyase activity and chalcone synthase activity were examined in roselle callus tissues incubated under different culture conditions. Phenylalanine ammonia-lyase activity was not affected by either the kind of auxin supplemented to the medium or light regime. In contrast, chalcone synthase activity was markedly suppressed when the callus was cultured with a medium containing indole-3-acetic acid instead of 2,4-dichlorophenoxyacetic acid (2,4-D) or in the dark. The results imply that in roselle callus cultures chalcone synthase plays a more important role in anthocyanin biosynthesis regulated by 2,4-D and light irradiation than phenylalanine ammonialyase.Abbreviations LS Linsmaier and Skoog - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - PAL phenylalanine ammonia-lyase - CHS chalcone synthase     

Alpha-amylase inhibitors from roselle (Hibiscus sabdariffa Linn.) tea

A roselle (Hibiscus sabdariffa Linn.) tea extract was found to have high inhibitory activity against porcine pancreatic alpha-amylase. Hibiscus acid and its 6-methyl ester were respectively isolated as active principles from the 50% methanol and acetone extracts of roselle tea. The activity of each isolate was compared to that of structurally related citric acid, a previously known inhibitor of fungal alpha-amylase.     

Kenaf (Hibiscus cannabinus), roselle (Hibiscus sabdariffa)

Uses of 16 species of Hibiscus, section Furcaria DC, are discussed, with particular reference to kenaf (H. cannabinus L.) and roselle (H. sabdariffa L.), the two most important species grown commercially as fiber plants. Other uses of this versatile group include use as ornamentals and employment of various plant parts as food, medicine, wood for musical instruments, and in superstitious rites. Comparative morphology of certain vegetative and floral parts of 11 species and seeds of 12 species is discussed. Reaction of plants of 11 species to root-knot nematodes is reported. Ratings varied from highly susceptible to highly resistant; kenaf was more susceptible to nematodes than the other 10 species. Suggested origins, distribution, and genetic relationships of kenaf, roselle, and related species are treated.     

玫瑰茄悬浮细胞合成花青素的光效应研究

光照对悬浮培养的玫瑰茄细胞生物量无影响。随着光照强度增大,玫瑰茄细胞合成花青素的量增加,光照强度３１．０ｗ／ｍ２为饱和光照强度,超过该强度,玫瑰茄细胞合成花青素的量不再进一步增加;可见光中蓝光（４２０～５３０ｎｍ）是促进玫瑰茄细胞合成花青素最有效单色光,光强为３０．０ｗ／ｍ２,接种量为０．２ｇ湿细胞的５０ｍｌ培养液经１６ｄ培养,花青素产量为８．９７ｍｇ／５０ｍｌ,高出相同光照强度全色光下的６．５３ｍｇ／ｍｌ;黄光和绿光分别有一定的促进作用。当黑暗下的培养时间不超过８ｄ,后期经过不少于８ｄ的光照可以诱导出和全程光照相当的花青素产量,分别为６．６４和６．７２ｍｇ／５０ｍｌ（总培养时间不少于１６ｄ）。当黑暗下培养时间超过１２ｄ,由于营养成分消耗,光照延长,花青素产量也无法提高,添加１０ｍｌ新鲜培养基再进行光诱导,花青素产量可以提高（６．７５ｍｇ／５０ｍｌ）。     

玫瑰茄培养细胞中花色苷积累的代谢调节

调整培养基组成是提高玫瑰茄（ＨｉｂｉｓｃｕｓｓａｂｄａｒｉｆｆａＬ．）培养细胞花色苷产量的有效途径之一。改变Ｂ５培养其中钙、锰、铜等离子的浓度,增加了培养细胞花色苷的积累量,其中又以锰离子最为有效。其浓度为６＊１０＾４ｍｏｌ／Ｌ时,花色苷磁量达０．３４１ｇ／Ｌ,是对照的２．８２倍。     

中间产物对玫瑰茄培养细胞合成花青苷的影响

用Ｂ５培养基悬浮培养产色素的玫瑰茄培养细胞,培养１３天时,花青苷产量最高,为０．２５ｇ／Ｌ。培养基中添加终浓度为１０＾－６ｍｏｌ／Ｌ的外源Ｌ－Ｐｈｅ能够显著地增加产色素细胞花青苷的积累量。浓度为１０＾－７ｍｏｌ／Ｌ的槲皮素,可使悬浮培养的玫瑰茄细胞花青苷产量提高１．３倍,无论是Ｌ－Ｐｈｅ还是槲皮素均不能启动不产色素的细胞系产花青苷。     

A new plant type in roselle (Hibiscus sabdariffa L.) and inheritance of two of its main distinguishing characteristics

A new type of roselle (Hibiscus sabdariffa L.) differing from other types in respect of certain morphological and physiological characters is described. Characteristic glandless sepal (gs) was monofactorial recessive to the normal glanded sepal (Gs). The normal ovoid (Co) shape of capsules was incompletely dominant over the round shape (co) in the new type. No linkage was observed between gs and co.     

单色光对玫瑰茄悬浮细胞合成花青素的影响

玫瑰茄悬浮细胞合成花青素受光调节。将不同的单色滤光膜覆盖在摇瓶表面,控制光照强度,判定了蓝光是促进玫瑰茄细胞产花青素的最有效单色光;红光和橙光无效;其它单色光随其波长接近蓝光,正效应增强。单色光对玫瑰茄细胞培养过程的其它参数,如ｐＨ、降糖、细胞生物量等影响不大。     

Assessment of molecular diversity and evolutionary relationship of kenaf (Hibiscus cannabinus L.), roselle (H. sabdariffa L.) and their wild relatives

Kenaf (Hibiscus cannabinus L.) and roselle (H. sabdariffa L.) are valuable fibre crop species with diverse end use. Phylogenetic relationship of 73 accessions of kenaf, roselle and their wild relatives from 15 countries was assessed using 44 inter-simple sequence repeat (ISSR) and jute (Corchorus olitorius L.) specific simple sequence repeats (SSR) markers. A total of 113 alleles were identified of which 61.95 % were polymorphic. Jute specific SSR markers exhibited high polymorphism and resolving power in kenaf, although ISSR markers exhibited higher resolving power than SSR markers. Number of polymorphic alleles varied from 1 to 5 for ISSR and 1 to 6 for SSR markers. Cultivated species exhibited higher allele polymorphism (57 %) than the wild species (35 %), but the improved cultivars exhibited lower genetic diversity compared to germplasm accessions. Accessions with common genetic lineage and geographical distribution clustered together. Indian kenaf varieties were distinct from cultivars bred in other countries and shared more genetic homology with African accessions. High genetic diversity was observed in the Indian (J = 0.35–0.74) and exotic kenaf germplasm collections (J = 0.38–0.79), suggesting kenaf might have been introduced in India from Africa through Central Asia during early domestication. Genetic similarity-based cluster analysis was in close accordance with taxonomic classification of Hibiscus.     

Comparative analysis of diversification and population structure of kenaf (Hibiscus cannabinus L.) and roselle (H. sabdariffa L.) using SSR and RGA (resistance gene analogue) markers

Multiple DNA marker systems and complementary analytical approaches are often useful in population genetic analysis and speciation of plants. We investigated population structure of kenaf (Hibiscus cannabinus) and roselle (H. sabdariffa) for gaining insight in evolution and geographic separation of these crop species using SSR and RGA (resistance gene analogues) markers through Bayesian clustering and principal coordinate analysis (PCoA) methods. Genotyping by 12 SSR and 16 RGA markers amplified a total of 172 loci in the study population. The RGA markers generated higher number of alleles per marker (8.2) as compared to SSR (3.4), but exhibited lower heterozygosity in the population. Genetic variance and heterozygosity in roselle population for both marker systems were lower than in kenaf. RGA markers revealed higher variation among populations. Bayesian structure as well as PCoA analysis using RGA marker revealed distinct cluster for roselle, while SSR-based classification revealed high admixture. Results indicate geographic isolation and natural selection for adaptive RGA alleles in kenaf. The Indian kenaf landraces were distinct from the exotic kenaf accessions, suggesting separate lineage formation by geographic separation. Possible origin and domestication of roselle in South India is proposed.     

X-ray irradiation of excised embryos of mesta (Hibiscus cannabinus L. and H. sabdariffa L.)

Summary Excised embryos of Hibiscus spp. were treated with 1 kR to 6 kR of X-ray. Results indicate that germination was unaffected at this level of employed doses in both species, which in turn implies that the factors responsible for inhibition of germination are not present in the embryo. LD₅₀ values differed between varieties and species. Early varieties of both species were more sensitive to radiation than late varieties. Strikingly similar effects were observed for the varieties with smaller embryos over those with larger ones. Allopolyploid H. sabdariffa (2n=72) was more susceptible than diploid H. cannabinus (2n=36).Differences in mutation frequency exist between species with different levels of ploidy and between varieties within the same species. Most of the HC mesta varieties yielded higher mutation frequencies than those of HS mesta. Optimal dose for triggering mutations in all varieties (except the chlorophyll mutation variety of HC mesta) of the two species lies within a narrow range of 1 kR to 2 kR. Cent per cent seedling abnormalities is concomitant to LD₅₀; nevertheless, optimum dose for mutation frequency is independent of LD₅₀. Hence, the response should be viewed in terms of respective genotype. The advantages of the embryo irradiation technique are mentioned.     

Fermentation studies on roselle (Hibiscus sabdariffa) calyx neutralized with trona (sodium sesquicarbonate) at different concentrations

The effect of trona at different concentrations on the fermentation of roselle (Hibiscus sabdariffa) calyx was studied. Mixed flora of bacteria and fungi were isolated and identified from the fermenting calyx. The isolates were Micrococcus luteus, Staphylococcus aureus, Echerichia coli, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and A. flavus. The addition of trona to the samples raised the initial pH which affected the terminal pH. There was an increase in microbial population with increases in the concentration of trona, especially for bacterial population which ranged from 5.18 to 5.59 log10 c.f.u./ml. The changes in total titratable acidity level were inconsistent. Fermentation enhanced the protein content of the samples but significantly decreased (P < 0.05) the content of antinutritional factors (phytic acid and tannin).     

高产花色苷玫瑰茄细胞系的筛选

花色苷在植物中呈现粉红、红、紫红、紫等颜色,可以用作食品、药品及化妆品的着色剂,亦有药用价值。作为食品添加剂,颜色较合成色素自然,且安全无毒性。早在１９８７年,Ｍｉｚukami^[1]就建议用植物细胞培养物生产花色苷类代替合成色素。所有的植物培养细胞都是异源性的。各细胞之间产花色苷的能力相差很大^[2].因为产花色苷的细胞系带有颜色标记,所以容易识别并通过肉眼选择即可获得高产花色苷的细胞系。筛选的方法很多,如平板饲喂法^[3]、小细胞团法^[4]、细胞块法^[5]、肉眼观察直接挑选法及细胞分栋器法^[6]等。高产系花色苷的含量可增加几倍到几十倍,而且产量稳定。本文采用平板法及小细胞团法筛选高产花色苷的玫瑰茄(Hibiscus sabdariffa L.)细胞系。     

Variation of morphological and phytochemical traits in Roselle (Hibiscus sabdariffa L.) genotypes under different planting dates

Roselle (Hibiscus sabdariffa L.) is recognized as a valuable food crop due to its nutraceutical potential, rich pigment content, and medicinal properties. However, there is limited information on planting dates and suitable genotypes for roselle cultivation in arid and semi-arid regions. The objective of this field study was to investigate the influence of five planting dates (March, April, May, June, and July) on morphological and phytochemical characteristics of different roselle genotypes. Among genotypes, eight (Jiroft, Dalgan, Bampoor, Iranshahr, Nikshahr, Roodbar, Saravan, and Qaleganj) were Iranian landraces, and HA and HS-24 were originated from Ghana and Bangladesh, respectively. Planting date significantly influenced the number of branches, bolls and seeds per plant, sepal fresh weight, calyx and biomass yields, and harvest index in tested roselle landraces. The greatest morphological growth, fresh sepal weight (50.6 g plant^?1) and calyx yield (1519 kg ha^?1) were observed with the roselle planted in early May. Moreover, amounts of chlorophyll, flavonoids and antioxidant remained higher in roselle when planted between April–May. The number of branches/plant was found to be an important determinant of calyx yield (r = 0.707) in roselle. Dalgan resulted in the greatest growth and yields within a comparatively shorter period of time compared to other tested landraces. Both principal component analysis (PCA) and cluster analysis also indicated that Dalgan landrace possessed most suitable morphological and phytochemical traits among tested landraces, and therefore, it could be planted adopted for extensive cultivation settings in the arid and semi-arid regions.     

Improved plant regeneration in callus cultures of Sorghum bicolor (L.) Moench

Sorghum bicolor is a recalcitrant species for tissue culture regeneration and genetic transformation. Browning of explants is one of the factors limiting organ and tissue cultures. To overcome this, callus tissue was initiated from the shoot tips of in vitro germinating seeds (S. bicolor cv. Róna 1), and then cultured on modified MS media (Murashige and Skoog in Physiol Plant 15:473–497, 1962). In the first experiment, we tested callus induction on several media supplemented with casein hydrolysate, polyvinylpyrrolidone, honey, and sucrose. The best callus induction was recorded for the medium with honey and sucrose (80.0%) and for control medium (79.8%). Shoot regeneration was tested on the MS medium with 6-benzylaminopurine (BAP) supplemented with honey and sucrose at a 1:1 ratio (by weight) or with sucrose only. The highest percentage of calluses regenerating shoots was noted for those induced on the medium with sucrose and honey—approx. four times higher when compared to the control. Rooted plantlets were acclimatized with a 92% survival rate. In the second experiment, we analyzed culture responses to various ways of honey application to the induction media: honey (autoclaved or filtered) in presence or absence of sucrose. Supplementation of the medium with fructose, glucose, and maltose at a proportion typical for honey was also investigated. The explant and callus survival rates were similar to those of the honey–sucrose combination in the first experiment. Only presence of both sucrose and honey in the induction medium improved the total regeneration rate to 37.9% over the control (18.8%). Sucrose and honey appear to act synergistically for shoot regeneration in callus cultures of sorghum.

     

Anthocyanin production in callus cultures ofDaucus carota as influenced by nutrient stress and osmoticum

Summary Daucus carota callus developed red pigments under the influence of indole-3 acetic acid and kinetin. Maximum yield of anthocyanin at the end of 3 weeks was 5.4% on dry weight basis. The callus subjected to phosphate and nitrate stress produced 7.2% and 8.5% anthocyanin respectively. Feeding of sucrose at 7.5% level resulted in production of 15% anthocyanin. Mannitol as osmoticum had positive influence on anthocyanin production.     

In vitro culture of callus tissues and cell suspensions from okra (Hibiscus esculentus L.) and cotton (gossypium hirsutum L.)

Summary Methods are described for starting and maintaining callus-tissue cultures of twoMalvaceae, okra (Hibiscus esculentus L.) and cotton (Gossypium hirsutum L.). Okra callus was slow to initiate, but once started it was easy to maintain, in contrast to cotton, which was difficult to initiate and grow. Different media were required to establish the two species. The inclusion of 5 mg per liter of ascorbic acid aided in reducing the formation of black pigments in cotton callus. Hypocotyls of sterile young okra seedlings and leaves of cotton plants were used to produce the callus tissue. Rapidly growing cell suspensions of okra and cotton were obtained in B5 medium.     

Anthocyanin production in selected cell lines of grape (Vitis vinifera L.)

Summary Anthocyanin production of two lines ofVitis vinifera cell cultures, i.e., 5.4 and 13.1, which were obtained from the same starting material after 20 and 37 mo. of clonal selection, respectively, was investigated. Cell suspension cultures of lines 5.4 and 13.1 maintained an anthocyanin content of 0.44 ± 0.15 and 1.02 ± 0.31 mg·g⁻¹ fresh weight during 50 and 32 weekly maintenance subcultures, respectively. Under anthocyanin-promoting culture conditions, both lines showed an enhancement of their anthocyanin level by approximately fourfold. While line 5.4 accumulated peonidin 3-glucoside and cyanidin 3-glucoside in decreasing order, line 13.1 accumulated primarily peonidin 3-p-coumaroylglucoside with lesser amounts of malvidin monoglucoside. Results show that while the anthocyanin content was improved during the course of repeated selections, the anthocyanin composition was modified markedly favoring the accumulation of more metabolically-advanced anthocyanins.