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Induction, Subculture Cycle, and Regeneration of Callus in Safed Musli (Chlorophytum borivilianum) using Different Types of Phytohormones

Nakasha JJ, Sinniah UR, Kemat N, Mallappa KS

Pharmacogn Mag, 2016 Jul;12(Suppl 4):S460-S464.
PMID: 27761075

BACKGROUND: Chlorophytum borivilianum is an industrially valued medicinal crop. Propagation through seeds is not feasible because of low germination percentage and long dormancy period. Therefore, callus culture and plant regeneration can be an alternative to improve this crop production. Also, callus can serve as an alternative source of bioactive compounds.
OBJECTIVE: To evaluate the effect of different phytohormones on callus induction, subculture cycle, and regeneration studies of callus in C. borivilianum.
MATERIALS AND METHODS: Young shoot buds of C. borivilianum were inoculated on Murashige and Skoog medium fortified with 3% sucrose and different concentrations (0, 1, 5, 10, and 15 mg/L) of either naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid or indole-3-acetic acid and callus induction was evaluated up to four subcultures cycles. Shoot regeneration from callus was studied on Murashige and Skoog media fortified with 6-benzylaminopurine andkinetin or thidiazuron at varied levels (0, 0.5, 1, 2, and 3 mg/L). Microshoots were rooted on Murashige and Skoog media supplemented with 1.0 mg/L indole-3-butyric acid and plantlets were acclimatized before transferred to the natural conditions.
RESULTS: Callus induction was better evidenced on Murashige and Skoog media containing 5 mg/L 2,4-dichlorophenoxyacetic acid up to fourth subculture. Callus differentiated into shoots on Murashige and Skoog media fortified with 6-benzylaminopurine or kinetin, whereas thidiazuron completely failed to regenerate shoots. Furthermore, microshoots rooted on 1.0 mg/L indole-3-butyric acid containing Murashige and Skoog media. The rooted plantlets were successfully acclimatized and established in soil with 88.3% survivability.
CONCLUSION: The type of auxins played an important role in inducing callus tissue from shoot bud explants of Safed musli. In future, this in vitro protocol could benefit in crop improvement programs and serve as a new source of bioactive compounds from Safed musli callus tissue for various therapeutic applications.
SUMMARY: Explants de-differentiated to form callus on Murashige and Skoog media containing 5 mg/L 2,4-D up to fourth subculture.Callus re-differentiated into shoots on Murashige and Skoog media fortified with 0.5 mg/L BAP.In vitro rooting of shoots was achieved on 1.0 mg/L IBA containing Murashige and Skoog media.The rooted plantlets were successfully acclimatized and established in soil with 88.3% survivability. Abbreviations used: MS: Murashige and Skoog, NAA: naphthalene acetic acid, 2,4-D: 2,4-dichlorophenoxyacetic acid, IAA: indole-3-acetic acid, BAP: 6-benzylaminopurine, Kn: Kinetin, TDZ: thidiazuron, IBA: indole-3-butyric acid, RCBD: Randomized Complete Block Design, DMRT: Duncan's Multiple Range Test.

Matched MeSH terms: Sucrose
Increased susceptibility of post-weaning rats on high-fat diet to metabolic syndrome

Cheng HS, Ton SH, Phang SCW, Tan JBL, Abdul Kadir K

J Adv Res, 2017 Nov;8(6):743-752.
PMID: 29062573 DOI: 10.1016/j.jare.2017.10.002

The present study aimed to examine the effects of the types of high-calorie diets (high-fat and high-fat-high-sucrose diets) and two different developmental stages (post-weaning and young adult) on the induction of metabolic syndrome. Male, post-weaning and adult (3- and 8-week old, respectively) Sprague Dawley rats were given control, high-fat (60% kcal), and high-fat-high-sucrose (60% kcal fat + 30% sucrose water) diets for eight weeks (n = 6 to 7 per group). Physical, biochemical, and transcriptional changes as well as liver histology were noted. Post-weaning rats had higher weight gain, abdominal fat mass, fasting glucose, high density lipoprotein cholesterol, faster hypertension onset, but lower circulating advanced glycation end products compared to adult rats. This is accompanied by upregulation of peroxisome proliferator-activated receptor (PPAR) α and γ in the liver and receptor for advanced glycation end products (RAGE) in the visceral adipose tissue. Post-weaning rats on high-fat diet manifested all phenotypes of metabolic syndrome and increased hepatic steatosis, which are linked to increased hepatic and adipocyte PPARγ expression. Adult rats on high-fat-high-sucrose diet merely became obese and hypertensive within the same treatment duration. Thus, it is more effective and less time-consuming to induce metabolic syndrome in male post-weaning rats with high-fat diet compared to young adult rats. As male rats were selectively included into the study, the results may not be generalisable to all post-weaning rats and further investigation on female rats is required.

Matched MeSH terms: Sucrose
In vitro tuberization of Chlorophytum Borivilianum Sant & Fern (Safed musli) as influenced by sucrose, CCC and culture systems

Farshad Ashraf M, Abd Aziz M, Abdul Kadir M, Stanslas J, Farokhian E

Plant Cell Physiol, 2013 Aug;54(8):1356-64.
PMID: 23749812 DOI: 10.1093/pcp/pct083

This study focuses on the establishment of in vitro tuberization of Chlorophytum borivilianum using solid and liquid culture systems. A high in vitro tuberization rate on solid and stationary liquid Murashige and Skoog media was observed in the presence of 60 g l⁻¹ sucrose with 950, 1,265 and 1,580 µM 2-chloroethyl-trimethylammonium chloride (CCC). Application of a higher sucrose concentration of 90 g l⁻¹ showed a negative interaction with CCC on in vitro tuber number and days to in vitro tuber induction. For economic feasibility, 950 µM CCC with 60 g l⁻¹ sucrose was chosen as the best combination for in vitro tuberization in both solid and stationary liquid media. For optimization of in vitro tuber production,a comparison between solid, stationary liquid and shake liquid culture was carried out. Liquid culture with shaking at 80 r.p.m. resulted in a >2.5-fold increase in in vitro tuber production compared with solid culture.

Matched MeSH terms: Sucrose/pharmacology*
Fulltext In vitro and in vivo effects of standardized extract and fractions of Phaleria macrocarpa fruits pericarp on lead carbohydrate digesting enzymes

Ali RB, Atangwho IJ, Kuar N, Ahmad M, Mahmud R, Asmawi MZ

BMC Complement Altern Med, 2013;13:39.
PMID: 23425283 DOI: 10.1186/1472-6882-13-39

One vital therapeutic approach for the treatment of type 2 diabetes mellitus is the use of agents that can decrease postprandial hyperglycaemia by inhibiting carbohydrate digesting enzymes. The present study investigated the effects of bioassay-guided extract and fractions of the dried fruit pericarp of Phaleria macrocarpa, a traditional anti-diabetic plant, on α-glucosidase and α-amylase, in a bid to understand their anti-diabetic mechanism, as well as their possible attenuation action on postprandial glucose increase.

Matched MeSH terms: Sucrose/metabolism
Fulltext In Vitro Responses of Plant Growth Factors on Growth, Yield, Phenolics Content and Antioxidant Activities of Clinacanthus nutans (Sabah Snake Grass)

Haida Z, Nakasha JJ, Hakiman M

Plants (Basel), 2020 Aug 14;9(8).
PMID: 32823824 DOI: 10.3390/plants9081030

Clinacanthus nutans, commonly known as Sabah snake grass, is one of the more important medicinal plants in Malaysia's herbal industry. C. nutans has gained the attention of medical practitioners due to its wide range of bioactive compounds responsible for various biological activities, such as anti-cancer, anti-venom and anti-viral activities. Due to its high pharmacological properties, the species has been overexploited to meet the demands of the pharmaceutical industry. The present study was conducted to establish a suitable in vitro culture procedure for the mass propagation of C. nutans. Murashige and Skoog (MS) basal medium, supplemented with different types of cytokinins, auxins, basal medium strength and sucrose concentrations, were tested. Based on the results, a full-strength MS basal medium supplemented with 12 µM 6-benzylaminopurine (BAP) and 30 g/L sucrose was recorded as the best outcome for all the parameters measured including the regeneration percentage, number of shoots, length of shoots, number of leaves and fresh weight of leaves. In the analysis of the phenolics content and antioxidant activities, tissue-cultured leaf extracts assayed at 100 °C exhibited the highest phenolic content and antioxidant activities. The propagation of C. nutans via a plant tissue culture technique was recorded to be able to produce high phenolic contents as well as exhibit high antioxidant activities.

Matched MeSH terms: Sucrose
Improved stability of live attenuated vaccine gdhA derivative Pasteurella multocida B:2 by freeze drying method for use as animal vaccine

Oslan SNH, Halim M, Ramle NA, Saad MZ, Tan JS, Kapri MR, et al.

Cryobiology, 2017 12;79:1-8.
PMID: 29037980 DOI: 10.1016/j.cryobiol.2017.10.004

The efficacy of attenuated strain of gdhA derivative Pasteurella multocida B:2 mutant as a live vaccine to control haemorrhagic septicaemia (HS) disease in cattle and buffaloes has been demonstrated. In order to use P. multocida B:2 mutant as a commercial product, it is essential to optimise its formulation for high viability and stability of the live cells. The effectiveness of freeze-drying process using different protective agent formulations for improving cells viability was explored. Sugar and nitrogen compounds were used as protective agents in freeze-drying and the capability of these compounds in maintaining the viability of mutant P. multocida B:2 during subsequent storage was investigated. A complete loss in viability of freeze-dried mutant P. multocida B:2 was monthly observed until 6-12 months of storage at -30 °C, 4 °C and 27 °C when nitrogen compound or no protective agent was added. Trehalose and sucrose showed significantly high survival rate of 93-95% immediately after freeze-drying and the viability was retained during the subsequent storage at -30 °C and 4 °C. A smooth cell surface without any cell-wall damage was observed for the cells formulated with trehalose under scanning electron micrograph. This study presented a freeze-drying process generating a dried live attenuated vaccine formulation with high stability for commercial applications.

Matched MeSH terms: Sucrose/metabolism*
Fulltext Impact of soil field water capacity on secondary metabolites, phenylalanine ammonia-lyase (PAL), maliondialdehyde (MDA) and photosynthetic responses of Malaysian kacip fatimah (Labisia pumila Benth)

Jaafar HZ, Ibrahim MH, Mohamad Fakri NF

Molecules, 2012 Jun 13;17(6):7305-22.
PMID: 22695235 DOI: 10.3390/molecules17067305

A randomized complete block design 2 × 4 experiment was designed and conducted for 15 weeks to characterize the relationships between production of total phenolics, flavonoid, anthocyanin, leaf gas exchange, total chlorophyll, phenylalanine ammonia-lyase (PAL) and malondialdehyde (MDA) activity in two varieties of Labisia pumila Benth, namely the var. alata and pumila, under four levels of evapotranspiration replacement (ER) (100%; well watered), (75%, moderate water stress), (50%; high water stress) and (25%; severe water stress). The production of total phenolics, flavonoids, anthocyanin, soluble sugar and relative leaf water content was affected by the interaction between varieties and SWC. As the ER levels decreased from 100% to 25%, the production of PAL and MDA activity increased steadily. At the highest (100%) ER L. pumila exhibited signiﬁcantly higher net photosynthesis, apparent quantum yield, maximum efficiency of photosystem II (f(v)/f(m)) and lower dark respiration rates compared to the other treatment. The production of total phenolics, flavonoids and anthocyanin was also found to be higher under high water stress (50% ER replacement) compared to severe water stress (25% ER). From this study, it was observed that as net photosynthesis, apparent quantum yield and chlorophyll content were downregulated under high water stress the production of total phenolics, flavonoids and anthocyanin were upregulated implying that the imposition of high water stress can enhance the medicinal properties of L. pumila Benth.

Matched MeSH terms: Sucrose/chemistry
Fulltext Impact of prebiotics on equol production from soymilk isoflavones by two Bifidobacterium species

Mustafa SE, Mustafa S, Ismail A, Abas F, Abd Manap MY, Ahmed Hamdi OA, et al.

Heliyon, 2020 Oct;6(10):e05298.
PMID: 33134584 DOI: 10.1016/j.heliyon.2020.e05298

The influence of commercial prebiotics (fructo-oligosaccharides and inulin) and sugars (glucose and sucrose) on enhancing equol production from soymilk isoflavones by Bifidobacterium longum BB536 and Bifidobacterium breve ATCC 15700 was evaluated in vitro. Sterilized soymilk was inoculated with each bacterial species at 37 °C for 48 h. The growth and β-glucosidase enzyme activity for the two Bifidobacterium species in soymilk throughout fermentation were assessed. The highest viable count for B. breve (8.75 log CFU/ml) was reached at 36 h and for B. longum (8.55 log CFU/ml) at 24 h. Both bacterial species displayed β-glucosidase activity. B. breve showed increased enzyme activity (4.126 U) at 36 h, while B. longum exhibited maximum activity (3.935 U) at 24 h of fermentation. Among the prebiotics screened for their effect in isoflavones transformation to equol, inulin delivered the highest effect on equol production. The co-culture of B. longum BB536 and B. breve ATCC15700 in soymilk supplemented with inulin produced the highest level (11.49 mmol/l) of equol at 48 h of fermentation process. Level of daidzin declined whereas that of daidzein increased, and then gradually decreased due to formation of equol when soymilk was fermented using bifidobacterial. This suggests that the nutritional value of soymilk may be increased by increasing bioavailability of the bioactive ingredients. Collectively these data identify probiotics and prebiotic combinations suitable for inclusion in soymilk to enhance equol production.

Matched MeSH terms: Sucrose
Impact of national income and inequality on sugar and caries relationship

Masood M, Masood Y, Newton T

Caries Res, 2012;46(6):581-8.
PMID: 23006794 DOI: 10.1159/000342170

The aim of this study was to examine the impact that national income and income inequality in high and low income countries have on the relationship between dental caries and sugar consumption.

Matched MeSH terms: Dietary Sucrose/adverse effects; Dietary Sucrose/supply & distribution*
Fulltext Health benefits of Stevia rebaudiana Bertoni as zero calorie natural sweetener: a review

Haida, Z., Asikin, A., Hakiman, M.

International Food Research Journal, 2020;27(5):783-789.
MyJurnal

Stevia rebaudiana Bertoni is a perennial herbal species with a number of therapeutic properties. The leaves of S. rebaudiana are the most important part of the plant as they contain high
level of sweetener compounds known as steviol glycosides; which are known to possess
antioxidant, antimicrobial, and antifungal activities. Among the steviol glycosides, stevioside
and rebaudioside A are the most abundant sweetening compounds of interest, documented to
be 30 - 250 folds sweeter than sucrose or refined sugar. The therapeutic properties of the
species make it more interesting as they play important roles as anti-hyperglycaemic agent to
lower glucose levels in the blood, thus commonly used in the treatment of diabetes mellitus
and obesity. Known to have sweet tasting leaves, its extract which contains antioxidant
compounds have been used to treat cancer, reduce inflammatory, and hypertension. S. rebaudiana has drawn the attention of health-conscious fitness lovers all over the world as non-caloric sweetener, and has been widely cultivated for being the most suitable sweetening substitution.

Matched MeSH terms: Sucrose
Fulltext Habitual sugar intake and cognitive impairment among multi-ethnic Malaysian older adults

Chong CP, Shahar S, Haron H, Din NC

Clin Interv Aging, 2019;14:1331-1342.
PMID: 31413554 DOI: 10.2147/CIA.S211534

Background: Sugar is widely consumed in Malaysia, and the excessive intake of sugar has been associated with cognitive functions. However, the association between sugar intake and cognitive impairment among Malaysian older adults is yet to be determined. Purpose: The objective of this study was to evaluate the associations between types and sources of sugar intake and cognitive functions and to identify their risk in predicting cognitive impairment (MMSE score <24). Subjects and methods: A total of 1,209 subjects aged ≥60 years were recruited through multi-stage random sampling from selected states in Malaysia. Dietary intake was derived using a 7-day dietary history questionnaire and supplemented with a quantitative food frequency questionnaire for added sugar intake. Results: The prevalence of cognitive impairment as defined by Mini-Mental State Examination (MMSE) less than 24 was 31.9%, while the prevalence of mild cognitive impairment was 13.1%. The median (IQR) for total sugar intake was 44.60 g/day (26.21-68.81) or 8 tsp, and free sugar intake was 33.08 g/day (17.48-57.26) or 6 tsp. The higher intake of total sugars, free sugars, sucrose, lactose, sugar-sweetened beverages, sugar-sweetened cakes, and dessert was found to be significantly associated with a lower MMSE score, after adjusting for covariates. On the other hand, the consumption of cooked dishes and fruits was significantly associated with a better MMSE score. The adjusted OR for risk of cognitive impairment (MMSE score <24) was 3.30 (95% CI 2.15-5.08) for total sugars and 3.58 (95% CI 2.32-5.52) for free sugars, comparing the highest with the lowest intake percentiles. Conclusion: Excessive sugar consumption among older adults showed a notable association with poor cognitive functions, but longitudinal studies and clinical trials are further needed to clarify the direction of causality and to investigate the underlying mechanism.

Matched MeSH terms: Dietary Sucrose/administration & dosage*
Fulltext Growth kinetics of citrus suhuiensis cell suspension

Noor Illi Mohamad Puad, Muhammad Alif Sarji, Nur Alia M. Fathil, Muhammad Yusuf Abduh

Biological and Natural Resources Engineering Journal, 2018;1(1):1-13.
MyJurnal

Citrus is one of the major commodities in many countries including Malaysia.
However, production of citrus including Citrus suhuiensis (C. suhuiensis) is declining due to
diseases and inability to withstand low temperatures. Plant cultures such as cell suspension have the
potential in propagating disease-free and healthy Citrus fruits with value-added characteristics.
However, studies related to C. suhuiensis is still scarce. Therefore, the growth kinetics of C.
suhuiensis cell suspension culture was studied. Friable callus of C. suhuiensis which was induced
from seeds was inoculated into MS medium with 30 g/L sucrose, 0.5 g/L malt extract and 2.0 mg/L
2, 4-D for the cell suspension initiation. Several batch experiments using a few types of sugars
(sucrose, glucose and fructose) were carried out. The cell dry weight (CDW) of C. suhuiensis was
recorded for 30 days of culture period and residual sugars in the medium were analyzed using
HPLC. Cells grown in 30 g/L sucrose achieved the highest CDW (9.559 g/L) with µmax equals to
0.00512/h, compared to glucose and fructose. In addition, sucrose is the preferred carbon source
with the highest uptake rate (0.213 g/L·h). Cells completely hydrolyzed sucrose into glucose and
fructose after 5 days of inoculation. All sugars were completely utilized by C. suhuiensis cells after
25 days. The kinetic growth parameters determined from batch experiments were then used for
model simulation and verification in MATHCAD 15. After adjustments and refinement to the
selected kinetic parameters, the model has fairly described and predicted the growth and sugars
profile of C. suhuiensis cells. The proposed model can be used to predict sucrose hydrolysis, glucose
and fructose formation from sucrose and their consumption by plant cells and also for larger scale
of growth.

Matched MeSH terms: Sucrose
Glycyrrhizic acid prevents high calorie diet-induced metabolic aberrations despite the suppression of peroxisome proliferator-activated receptor γ expression

Cheng HS, Yaw HP, Ton SH, Choy SM, Kong JM, Abdul Kadir K

Nutrition, 2016 Sep;32(9):995-1001.
PMID: 27130470 DOI: 10.1016/j.nut.2016.02.002

OBJECTIVE: To investigate the effects of glycyrrhizic acid supplementation on glucose and lipid metabolism in rodents consuming a high-fat, high-sucrose diet.
METHODS: Twenty-four male, 8-week old Sprague Dawley rats with an initial weight of 160 to 200 g were randomised into three groups (n = 6 for each group): groups A (standard rat chow), B (high-fat, high-sucrose diet), and C (high-fat, high-sucrose diet + 100 mg/kg/d of glycyrrhizic acid via oral administration). The rats were treated accordingly for 4 wk. Glycaemic parameters, lipid profile, stress hormones, and adiponectin levels were measured after the treatment. Relative gene expressions of peroxisome proliferator-activated receptor α and γ, lipoprotein lipase as well as gluconeogenic enzymatic activities in different tissues were also determined.
RESULTS: Consumption of high-fat, high-sucrose diet triggered hyperglycaemia, insulin resistance, and dyslipidemia, which were effectively attenuated by supplementation with glycyrrhizic acid. Glycyrrhizic acid supplementation also effectively reduced circulating adrenaline, alleviated gluconeogenic enzymes overactivity, and promoted the upregulation of lipoprotein lipase expression in the cardiomyocytes and skeletal muscles. A high calorie diet also triggered hypoadiponectinaemia and suppression of peroxisome proliferator-activated receptor γ expression, which did not improve with glycyrrhizic acid treatment.
CONCLUSION: Supplementation with glycyrrhizic acid could alleviate high calorie diet-induced glucose and lipid metabolic dysregulations by reducing circulatory stress hormones, normalizing gluconeogenic enzyme activities, and elevating muscular lipid uptake. The beneficial effects of these bioactivities outweighed the adverse effects caused by diet-induced repression of peroxisome proliferator-activated receptor γ expression, resulting in the maintenance of lipid and glucose homeostasis.

Matched MeSH terms: Sucrose/administration & dosage*
Fulltext Glycyrrhizic acid can attenuate metabolic deviations caused by a high-sucrose diet without causing water retention in male Sprague-Dawley rats

Fernando HA, Chandramouli C, Rosli D, Lam YL, Yong ST, Yaw HP, et al.

Nutrients, 2014 Nov 04;6(11):4856-71.
PMID: 25375630 DOI: 10.3390/nu6114856

Glycyrrhizic acid (GA) ameliorates many components of the metabolic syndrome, but its potential therapeutic use is marred by edema caused by inhibition of renal 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2). We assessed whether 100 mg/kg per day GA administered orally could promote metabolic benefits without causing edema in rats fed on a high-sucrose diet. Groups of eight male rats were fed on one of three diets for 28 days: normal diet, a high-sucrose diet, or a high-sucrose diet supplemented with GA. Rats were then culled and renal 11β-HSD2 activity, as well as serum sodium, potassium, angiotensin II and leptin levels were determined. Histological analyses were performed to assess changes in adipocyte size in visceral and subcutaneous depots, as well as hepatic and renal tissue morphology. This dosing paradigm of GA attenuated the increases in serum leptin levels and visceral, but not subcutaneous adipocyte size caused by the high-sucrose diet. Although GA decreased renal 11β-HSD2 activity, it did not affect serum electrolyte or angiotensin II levels, indicating no onset of edema. Furthermore, there were no apparent morphological changes in the liver or kidney, indicating no toxicity. In conclusion, it is possible to reap metabolic benefits of GA without edema using the current dosage and treatment time.

Matched MeSH terms: Dietary Sucrose/administration & dosage*; Dietary Sucrose/adverse effects
Fruit characteristics, soluble sugar compositions and transcriptome analysis during the development of Citrus maxima "seedless", and identification of SUS and INV genes involved in sucrose degradation

Deng S, Mai Y, Niu J

Gene, 2019 Mar 20;689:131-140.
PMID: 30576805 DOI: 10.1016/j.gene.2018.12.016

Citrus maxima "seedless" is originally from Malaysia, and now is widely cultivated in Hainan province, China. The essential features of this cultivar are thin skin, green epicarp and seedless at the ripening stage. Here, using C. maxima "seedless" as experimental material, we investigated the physical and inclusion indicators, and found the accumulation of storage compounds during 120-210 DAF leading to inconsistent increase between volume and weight. Component analysis of soluble sugar indicated that arabinose and xylose have a high content in early development of pummelo juice sacs (PJS), whereas fructose, glucose and sucrose show a significant increase during PJS maturation. To clarify a global overview of the gene expressing profiles, the PJSs from four periods (60, 120, 180 and 240 DAF) were selected for comparative transcriptome analysis. The resulting 8275 unigenes showed differential expression during PJS development. Also, the stability of 11 housekeeping genes were evaluated by geNorm method, resulting in a set of five genes (UBC, ACT, OR23, DWA2 and CYP21D) used as control for normalization of gene expression. Based on transcriptome data, 5 sucrose synthases (SUSs) and 10 invertases (INVs) were identified to be involved in sucrose degradation. Importantly, SUS4 may be responsible for arabinose and xylose biosynthesis to form the cell wall in early development, while SUS3 and VIN2 may be important in the accumulation of soluble hexose leading to cell expansion through an osmotic-independent pathway in late development. The information provides valuable metabolite and genetic resources in C. maxima "seedless", and is important for achieving high fruit yield and quality.

Matched MeSH terms: Sucrose/metabolism*
First Report of a Soft Rot of Phalaenopsis aphrodita Caused by Dickeya dieffenbachiae in China

Zhou JN, Lin BR, Shen HF, Pu XM, Chen ZN, Feng JJ

Plant Dis, 2012 May;96(5):760.
PMID: 30727539 DOI: 10.1094/PDIS-11-11-0942

Phalaenopsis orchids, originally from tropical Asia, are mainly planted in Thailand, Singapore, Malaysia, the Philippines, and Taiwan and have gained popularity from consumers all over the world. The cultivation area of Phalaenopsis orchids has been rising and large-scale bases have been established in mainland China, especially South China because of suitable environmental conditions. In September 2011, a soft rot of Phalaenopsis aphrodita was found in a Phalaenopsis planting base in Guangzhou with an incidence of ~15%. Infected plants initially showed water-soaked, pale-to-dark brown pinpoint spots on leaves that were sometimes surrounded by a yellow halo. Spots expanded rapidly with rising humidity and temperatures, and in a few days, severely extended over the blade with a light tan color and darker brown border. Lesions decayed with odorous fumes and tissues collapsed with inclusions exuding. The bacterium advanced to the stem and pedicle. Finally, leaves became papery dry and the pedicles lodged. Six diseased samples were collected, and bacteria were isolated from the edge of symptomatic tissues after sterilization in 0.3% NaOCl for 10 min, rinsing in sterile water three times, and placing on nutrient agar for culture. Twelve representative isolates were selected for further characterization. All strains were gram negative, grew at 37°C, were positive for indole production, and utilized malonate, glucose, and sucrose but not glucopyranoside, trehalose, or palatinose. Biolog identification (version 4.20.05, Hayward, CA) was performed and Pectobacterium chrysanthemi (SIM 0.868) was confirmed for the tested isolates (transfer to genus Dickeya). PCR was used to amplify the 16S rDNAgene with primers 27f and 1492r, dnaX gene with primers dnaXf and dnaXr (3), and gyrB gene with primers gyrBf (5'-GAAGGYAAAVTKCATCGTCAGG-3') and gyrB-r1 (5'-TCARATATCRATATTCGCYGCTTTC-3') designed on the basis of the published gyrB gene sequences of genus Dickeya. BLASTn was performed online, and phylogeny trees (100% bootstrap values) were created by means of MEGA 5.05 for these gene sequences, respectively. Results commonly showed that the representative tested strain, PA1, was most homologous to Dickeya dieffenbachiae with 98% identity for 16S rDNA(JN940859), 97% for dnaX (JN989971), and 96% for gyrB (JN971031). Thus, we recommend calling this isolate D. dieffenbachiae PA1. Pathogenicity tests were conducted by injecting 10 P. aphrodita seedlings with 100 μl of the bacterial suspension (1 × 108 CFU/ml) and another 10 were injected with 100 μl of sterile water as controls. Plants were inoculated in a greenhouse at 28 to 32°C and 90% relative humidity. Soft rot symptoms were observed after 2 days on the inoculated plants, but not on the control ones. The bacterium was isolated from the lesions and demonstrated identity to the inoculated plant by the 16S rDNA sequence comparison. Previously, similar diseases of P. amabilis were reported in Tangshan, Jiangsu, Zhejiang, and Wuhan and causal agents were identified as Erwinia spp. (2), Pseudomonas grimontii (1), E. chrysanthemi, and E. carotovora subsp. carovora (4). To our knowledge, this is the first report of D. dieffenbachiae causing soft rot disease on P. aphrodita in China. References: (1) X. L. Chu and B. Yang. Acta Phytopathol. Sin. 40:90, 2010. (2) Y. M. Li et al. J. Beijing Agric. Coll. 19:41, 2004. (3) M. Sławiak et al. Eur. J. Plant Pathol. 125:245, 2009. (4) Z. Y. Wu et al. J. Zhejiang For. Coll. 27:635, 2010.

Matched MeSH terms: Sucrose
First Report of Pantoea anthophila Causing Soft Rot Disease in Clausena lansium (Wampee) in China

Zhou JN, Liu SY, Chen YF, Liao LS

Plant Dis, 2015 Mar;99(3):416.
PMID: 30699721 DOI: 10.1094/PDIS-10-14-1025-PDN

Clausena lansium, also known as wampee (Clausena wampi), is a plant species native to China, Vietnam, the Philippines, Malaysia, and Indonesia, where it is widely cultivated, and also grown in India, Sri Lanka, Queensland, Florida, and Hawaii, but less frequently (3). The fruit can be consumed fresh or made into juice, jam, or succade. In summer to fall 2014, a soft rot disease was found in a wampee planting region in Yunan County, Guangdong Province, China. On Sept. 18, we collected diseased samples from a wampee orchard with about 20% disease incidence. The infected fruit initially showed pinpoint spots on the peel, water-soaked lesions, and light to dark brown discoloration. Spots expanded in 2 days, and tissues collapsed after 5 days. Severely affected fruit showed cracking or nonodorous decay. Five diseased samples were collected, and causal agents were isolated from symptomatic tissues 1 cm under the peel after surface sterilization in 0.3% NaOCl for 10 min and rinsing in sterile water three times. Tissues were placed on a Luria Bertani (LB) plate for culture. Ten representative isolates were selected for further characterization. No colony was isolated from healthy tissues. Colonies were round, smooth, with irregular edges, and produced a yellow pigment in culture. Biolog identification (Version 4.20.05) showed that all strains were gram negative, negative for indole production, and utilized glucose, maltose, trehalose, sucrose, D-lactose, and pectin but not sorbitol or gelatin. The isolates were identified as Pantoea agglomerans (SIM 0.69). Multilocus sequence analysis (MLSA) was conducted for rapid classification of the strains. Sequences of atpD, gyrB, infB, and rpoB were amplified using corresponding primers (2). All sequences of the 10 isolates were identical in each gene. BLASTn was performed, and maximum likelihood trees based on the concatenated nucleotide sequences of the four genes were constructed using MEGA6. Bootstrap values after 1,000 replicates were expressed as percentages. Results showed that the tested strain named CL1 was most homologous to P. anthophila, with 98% identity for atpD (KM521543), 100% for gyrB (KM521544), infB (KM521545), and rpoB (KM521546). The 16S rRNA sequence (KM521542) amplified by primers 27f and 1492r shared 99% identity with that of P. anthophila M19_2C (JN644500). P. anthophila was previously reclassified from P. agglomerans (3); therefore, we suggest naming this wampee pathogen P. anthophila. Subsequently, 10 wampee fruits were injected with 20 μl of bacterial suspension (1 × 108 CFU/ml) of strains CL1 and CL2, respectively, and another 10 were injected with 20 μl of LB medium as controls, all kept at 28°C for 4 days. Symptoms similar to those of natural infections were observed on inoculated fruits but not on the negative controls. Bacteria were isolated from diseased tissues and further identified as P. anthophila by gyrB sequencing. P. anthophila was reported to naturally infect balsam and marigold (1,2). To our knowledge, this is the first report of P. anthophila naturally causing soft rot disease and cracking on C. lansium (wampee). References: (1) C. Brady et al. Syst. Appl. Microbiol. 31:447, 2008. (2) C. Brady et al. Int. J. Syst. Evol. Microbiol. 59:2339, 2009. (3) J. Morton. Fruits of Warm Climates. Echo Point Books & Media, Miami, FL, 1987.

Matched MeSH terms: Sucrose
Fulltext Fermentation of black tea broth (Kombucha): I. effects of sucrose concentration and fermentation time on the yield of microbial cellulose

Goh, W.N., Rosma, A., Kaur, B., Fazilah, A., Karim, A.A., Rajeev Bhat

International Food Research Journal, 2012;19(1):109-117.
MyJurnal

The yield and properties of cellulose produced from bacterial fermentation of black tea broth (known as Kombucha) were investigated in this study. The tea broth was fermented naturally over a period of up to 8 days in the presence of sucrose. Tea broth with a sucrose concentration of 90 g/l produced highest yield of bacterial cellulose (66.9%). The thickness and yield of bacterial cellulose increased with fermentation time. The bacterial cellulose production increased correspondingly with increased surface area:depth ratio. Changes in pH were related to the symbiotic metabolic activities of yeasts and acetic acid bacteria, and the counts of both of these in the tea broths were relatively higher than those in the cellulose layer. Findings from this study suggest that the yield of cellulose depends on many factors that need to be optimized to achieve maximum yield.

Matched MeSH terms: Sucrose
Fulltext Explorative Xylitol Detection from Lignocellulosic Biomass Fermentation of Coconut Husk Using HPLC C18 Column

Mariam-Aisha Fatima, Muhammad Fahmi Mehdin, Nurain Nasrudin, Neelam Shahab

Journal of Management & Science, 2018;16(2):29-38.
MyJurnal

Lignocellulosic biomass, found in wooden plant husks is a potent renewable material source which can be utilised to form various chemicals and biomaterials including polyols such as xylitol. Xylitol has been used commercially as an alternative to sucrose in many products as bulk sweetener in non-cariogenic confectionery as well as in diabetic diets and solutions for parenteral nutrition. Therefore, this study aims to optimise separation parameters of the Shimadzu high performance liquid chromatography (HPLC) (Model No: LC-20A) and quantify the potential of coconut husk as substrate for Candida albicans in producing xylitol using HPLC. Pretreatment to depolymerise components of biomass i.e. cellulose, hemicellulose and lignin were done using dilute acid hydrolysis method which yielded fermentable reducing sugars, xylose. Xylose is a monosaccharide with an aldehyde functional group, a reducing sugar which is then utilised and fermented by the yeast Candida albicans to form xylitol. In this study, the media used for fermenting pretreated coconut husk and Candida albicans is a synthetic defined (SD) minimal broth. Growth curves against concentration of reducing sugar were plotted to determine utilisation and production trends with specific mixes of carbon sources. The presence of reducing sugars were tested using 3,5-dinitrosalicylic acid (DNS) assay in pretreated coconut husk yielded 13.22 g/L. In order to analyse the product with HPLC, an existing protocol is modified for Hypersil GOLD™ C18 column with acetonitrile as the mobile phase. Results obtained from HPLC analyses using developed protocol suggested the formation of xylitol from the fermentation of pretreated husk by Candida albicans. These can be followed by purification of fermented media in obtaining a better separation of contaminating peaks.

Matched MeSH terms: Sucrose
Fulltext Experimental mixture design as a tool to optimize the growth of various Ganoderma species cultivated on media with different sugars

Goh YK, Marzuki NF, Tan SY, Tan SS, Tung HJ, Goh YK, et al.

Mycology, 2016;7(1):36-44.
PMID: 30123614 DOI: 10.1080/21501203.2015.1137985

The influence of different medium components (glucose, sucrose, and fructose) on the growth of different Ganoderma isolates and species was investigated using mixture design. Ten sugar combinations based on three simple sugars were generated with two different concentrations, namely 3.3% and 16.7%, which represented low and high sugar levels, respectively. The media were adjusted to either pH 5 or 8. Ganoderma isolates (two G. boninense from oil palm, one Ganoderma species from coconut palm, G. lingzhi, and G. australe from tower tree) grew faster at pH 8. Ganoderma lingzhi proliferated at the slowest rate compared to all other tested Ganoderma species in all the media studied. However, G. boninense isolates grew the fastest. Different Ganoderma species were found to have different sugar preferences. This study illustrated that the mixture design can be used to determine the optimal combinations of sugar or other nutrient/chemical components of media for fungal growth.

Matched MeSH terms: Sucrose

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