Multiple metabolite profiles uncover remarkable bioactive compounds and metabolic characteristics of noni fruit (Morinda citrifolia L.) at various stages of ripeness.

This study aimed to investigate the changes in physicochemical properties, bioactive compounds, and metabolic characteristics of noni fruit at different ripeness levels. The results showed that there were significant differences in physicochemical properties. HPLC analysis was conducted, revealing succinic acid, scopoletin, deacetylasperulosidic acid, and asperulosidic acid were key bioactive compounds as the fruit ripened. Additionally, 4 differentbiomarkers (isocitric acid, 4,4-thiodiphenol, lobaric acid, and octocrylene), identified using 1HNMR and LC-IT-TOF-MS, were found to have a VIP value over 1. The results from HS-GC-IMS demonstrated noteworthy that 14 volatile compounds were identified as highly discriminative features during fruit ripening. Furthermore, correlation analysis showed that different ripeness had significant effects on bioactive components and functional activities, e.g., the inhibition rate of enzyme and E. coli of noni fruit with different ripeness exceeded 90% at the last stage. This study contributes new insights into the effective utilization of bioactive ingredients in noni fruit.

Characterization of Chemical Component Variations in Different Growth Years and Tissues of Morindae Officinalis Radix by Integrating Metabolomics and Glycomics.

Morindae Officinalis Radix (MOR), the dried root of Morinda officinalis F.C. How (Rubiaceae), is a popular food supplement in southeastern China for bone protection, andrological, and gynecological healthcare. In clinical use, 3-4 year old MOR is commonly used and the xylem is sometimes removed. However, there is no scientific rationale for these practices so far. In this study, metabolomics and glycomics were integrated using multiple chromatographic and mass spectrometric techniques coupled with multivariate statistical analysis to investigate the qualitative and quantitative variations of secondary metabolome and glycome in different growth years (1-7 years) and tissues (xylem and cortex) of MOR. The results showed that various types of bioactive components reached a maximum between 3 and 4 years of growth and that the xylem contained more potentially toxic constituents but less bioactive components than the cortex. This study provides the chemical basis for the common practice of using 3-4 year old MOR with the xylem removed.

Biochemistry and Cell Wall Changes Associated with Noni (Morinda citrifolia L.) Fruit Ripening.

Quality and compositional changes were determined in noni fruit harvested at five ripening stages, from dark-green to thaslucent-grayish. Fruit ripening was accompanied by acidity and soluble solids accumulation but pH diminution, whereas the softening profile presented three differential steps named early (no significant softening), intermediate (significant softening), and final (dramatic softening). At early step the extensive depolymerization of hydrosoluble pectins and the significantly increment of pectinase activities did not correlate with the slight reduction in firmness. The intermediate step showed an increment of pectinases and hemicellulases activities. The final step was accompanied by the most significant reduction in the yield of alcohol-insoluble solids as well as in the composition of uronic acids and neutral sugars; pectinases increased their activity and depolymerization of hemicellulosic fractions occurred. Noni ripening is a process conducted by the coordinated action of pectinases and hemicellulases that promote the differential dissasembly of cell wall polymers.

Água salina e biofertilizante bovino na produção de frutos e alocação de biomassa em noni (Morinda citrifolia L. ) / Saline water and bovine biofertilizer on yield and biomass allocation in noni plants (Morinda citrifolia L. )

RESUMO: Têm sido evidenciado que os compostos orgânicos exercem ação mitigadora dos efeitos tóxicos dos sais às plantas, promovendo maior crescimento e desenvolvimento em solos com excesso de sais. Nessa direção, um experimento foi desenvolvido para avaliar os efeitos do biofertilizante bovino na produção de frutos e alocação de biomassa pelos diversos órgãos de plantas de noni, irrigadas com águas salinas. Os tratamentos foram distribuídos em blocos ao acaso, com quatro repetições e duas plantas por parcela, adotando o arranjo fatorial 5 x 2, correspondente a salinidade das águas de irrigação de 0,5; 1,5; 3,0; 4,5; 6,0 dS m-1, em substratos sem e com biofertilizante bovino, aplicado uma única vez, em volume correspondente a 10% do volume do substrato, um dia antes da instalação do experimento. As variáveis avaliadas foram: salinidade do solo, expressa pela condutividade elétrica do extrato de saturação do solo; número de frutos; massa média de fruto; produção de frutos por planta; e alocação de biomassa pelas raízes, caules, folhas e frutos. O aumento da salinidade da água de irrigação elevou o caráter salino do solo desde "não salino" para "fortemente salino" e, inibiu a produção de frutos e a acumulação de biomassa seca das plantas de noni em geral, mas, com maior intensidade nos tratamentos sem o insumo orgânico. O biofertilizante estimula o crescimento e a produção de plantas de noni cultivadas sob irrigação com águas de alta salinidade.

ABSTRACT: Organic compounds have shown to exercise a mitigating action on salts in plants and they promote growth and development in salt stress environments. In this sense, an experiment was carried out in order to evaluate the effects of saline water irrigation and bovine biofertilizer on yield and biomass allocation by the various organs of noni plants. Treatments were arranged in a randomized block design with four replications and two plants per plot using the 5 x 2 factorial design, which correspond to the salinity levels of irrigation water of 0.5, 1.5, 3.0, 4.5 and 6.0 dS m-1 in substrates with and without bovine biofertilizer, applied to the soil once, in the volume corresponding to 10% of the volume of the substrate one day before the implementation of the experiment. The variables evaluated were soil salinity, expressed as electrical conductivity of the soil saturation extract, fruit number, fruit weight and fruit yield per plant, and biomass allocation by roots, stems, leaves and fruit. The increasing salinity of the water for irrigation increased the saline character of the soil from non-saline to saline soil and inhibited the production and accumulation of dry biomass in noni plants in general but with more intensity in the plants of the treatments without bovine biofertilizer applied to the soil in liquid form. The biofertilizer stimulates the growth and production of noni plants grown under irrigation with high salinity water.

Desenvolvimento vegetativo do noni (Morindacitrifolia L. ) sob diferentes sistemas e lâminas de irrigação / Vegetative development of noni (Morinda citrifolia L. ) under different irrigation systems and water depths

O noni é uma cultura de origem asiática que vem despertando interesse de agricultores brasileiros em virtude de suas propriedades fitoterápicas na medicina popular e de seu elevado valor comercial; contudo, são escassas as informações quanto ao manejo da cultura em geral e inexistentes estudos a respeito da irrigação da cultura no Brasil. Objetivou-se com este estudo definir, entre dois sistemas de irrigação e cinco laminas d´água, qual a melhor condição para o desenvolvimento vegetativo da cultura. O experimento foi realizado no período de maio de 2009 a março de 2010, em Trairi-CE. Utilizou-se o delineamento experimental de blocos casualizados e dois sistemas de irrigação localizada (gotejamento e microaspersão) dispostos em faixas verticais. Dentro de cada faixa foram distribuídos os blocos com as parcelas dos cincos níveis de irrigação correspondentes à aplicação de frações da evapotranspiração de referência FAO (ETo) de 40%, 60%, 80% 100% e 120%. Para o sistema de irrigação por microaspersão as lâminas correspondentes foram: 109,3 mm, 135,8 mm, 187,9 mm, 227,8 mm e 297,7 mm e para sistema de irrigação por gotejamento: 83,4 mm, 1237 mm, 166,5 mm, 204,7 mm e 254,4 mm, respectivamente. Foram utilizadas cinco repetições. O desenvolvimento vegetativo de plantas de noni foi influenciado tanto pelos sistemas de irrigação, quanto pelas lâminas d´água aplicadas. A lâmina de irrigação equivalente a 227,8 mm, correspondente a 100% da ETo, na irrigação por microaspersão, proporcionou maiores ganhos de diâmetro do coleto, altura da planta, e diâmetro de copa.

Noni is a crop of Asian origin that has piqued the interest of Brazilian farmers because of its properties in herbal folk medicine and its high commercial value. However, there is little information about the crop management in general, and particularly on the irrigation of the crop in Brazil. The objective of this study was to define what localized irrigation system (drip or micro sprinklers) and water depth that provide the best vegetative growth of noni plants. The experiment was conducted from May 2009 to March 2010 in the city of Trairi, Brazil. We used a split block experimental design, with two irrigation systems (drip and micro sprinklers) arranged in vertical lines. Within each line, five irrigation levels, corresponding to the application of fractions of the reference evapotranspiration (ETo) 40% 60% 80% 100% and 120%, were distributed in five blocks. For the micro sprinkler irrigation system, the corresponding slides were: 109.3 mm, 135.8 mm, 187.9 mm, 227.8 mm and 297.7 mm, respectively, and for the drip irrigation system, they were: 83.4 mm, 123.7 mm, 166.5 mm, 204.7 mm and 254.4 mm, respectively. The noni vegetative growth was influenced both by the irrigation systems and the depths of irrigation applied. The application of irrigation depths equivalent to 227.8 mm, which corresponds to 100% of ETo with the use of micro sprinklers, provided the largest gains in stem diameter, plant height and canopy diameter.

Transformation of Morinda citrifolia via simple mature seed imbibition method.

Morinda citrifolia, is a valuable medicinal plant with a wide range of therapeutic properties and extensive transformation study on this plant has yet been known. Present study was conducted to establish a simple and reliable transformation protocol for M. citrifolia utilising Agrobacterium tumefaciens via direct seed exposure. In this study, the seeds were processed by tips clipping and dried and subsequently incubated in inoculation medium. Four different parameters during the incubation such as incubation period, bacterial density, temperature and binary vectors harbouring beta-glucuronidase (GUS) gene (pBI121 and pGSA1131), were tested to examine its effect on transformation efficiency. The leaves from the treated and germinated seedlings were analysed via Polymerase Chain Reaction (PCR), histochemical assay of the GUS gene and reverse transcription-PCR (RT-PCR). Results of the study showed that Agrobacterium strain LBA4404 with optical density of 1.0 and 2 h incubation period were optimum for M. citrifolia transformation. It was found that various co-cultivation temperatures tested and type of vector used did not affect the transformation efficiency. The highest transformation efficiency for M. citrifolia direct seed transformation harbouring pBI121 and pGSA1131 was determined to be 96.8% with 2 h co-cultivation treatment and 80.4% when using bacterial density of 1.0, respectively. The transformation method can be applied for future characterization study of M. citrifolia.

[Tissue culture and polyploidy induction of Morinda officinalis].

OBJECTIVE: To establish an effective protocol for plant generation and induce polyploidy of Morinda offcinalis. METHOD: Callus was induced from immature embryo of M. officinalis and polyploidy was inducted by using colchicine treatment method. Chromosome was detected by flow cytometry. RESULT AND CONCLUSION: The highest induction rate of polyploidy was 18.40%, which was obtained with 500 mg x L(-1) colchicine treatment for 5 days. Roots of polyploid were bigger than diploid. Advantages of using immature embryo as explants are easy for sterilization, higher rate of callus induction and low degree dedifferentiation. The induced polyploidy of M. officinalis may have a value for spread of cultivation.

Desponte e embebição em sementes de noni (Morinda citrifolia L. ) / Lopping and soaking in seeds of noni (Morinda citrifolia L. )

O trabalho teve como objetivo testar a germinação em sementes de noni sob diferentes tempos de embebição, com e sem o desponte das sementes. Para isso, instalou-se um experimento em casa de vegetação no delineamento inteiramente casualizado com 10 tratamentos e 4 repetições de 25 sementes , sendo constituídos por: 1. Testemunha; 2. Sementes com desponte do lado oposto ao embrião; 3. Sementes intactas com 12 horas de embebição; 4. Sementes intactas com 24 horas de embebição; 5. Sementes intactas com 36 horas de embebição; 6. Sementes intactas com 48 horas de embebição; 7. Sementes com desponte com 12 horas de embebição; 8. Sementes com desponte com 24 horas de embebição; 9. Sementes com desponte com 36 horas de embebição e 10. Sementes com desponte com 48 horas de embebição. As características avaliadas foram: porcentagem de emergência, Índice de velocidade de emergência, altura de plântulas, comprimento de raiz e peso de massa seca das plântulas aos 46 dias após a instalação do experimento. Houve diferença significativa entre os diferentes tempos de embebição e o tratamento 10 (Desponte com 48 hs de embebição) que obteve os melhores resultados para a percentagem de emergência, índice e velocidade de emergência e peso seco. Quanto a altura de plântula, o tratamento 1 (Testemunha) foi o maior, enquanto no tratamento 2 (desponte) se verificou o maior comprimento de raiz. Embora não tenha diferido da testemunha, o melhor tratamento foi o uso do desponte + 48 horas de embebição.

The study aimed to test the germination of seeds of noni under different soaking times, with and without the seed emerges. For this, an experiment was installed in a greenhouse in a completely randomized design with 10 treatments and 4 replicates of 25 seeds, being composed of: 1. Witness 2. Seeds with the blunt side opposite the embryo 3. Intact seeds with 12 hours of soaking, 4. Intact seeds with 24 hours of soaking, 5. Intact seeds with 36 hours of soaking, 6. Intact seeds with 48 hours of soaking; 7. Seeds with cutting with 12 hours of soaking; 8. Seeds with cutting with 24 hours of soaking; 9. Seeds with cutting with 36 hours of soaking and 10. Seeds with cutting with 48 hours of soaking. The characteristics evaluated were: percentage of emergence, speed of emergence index, seedling height, root length and weight of the dry mass of seedlings 46 days after the experiment. There were significant differences between the different times of soaking and treatment 10 (cutting with 48 hours of soaking) who obtained the best results for the percentage of emergency and emergency speed index and dry weight. The height of the seedling, one treatment (control) was the highest, while in treatment 2 (blunt) registered the largest root length. Although the witness did not differ, the best treatment was the use of blunt + 48 hours of soaking.

Medium salt strength induced changes in growth, physiology and secondary metabolite content in adventitious roots of Morinda citrifolia: the role of antioxidant enzymes and phenylalanine ammonia lyase.

In an attempt to improve growth and secondary metabolite production, and to understand the possible mechanism involved in relation to the changes in physiology and activities of antioxidant enzymes, we cultured Morinda citrifolia adventitious roots in different strength (0.25, 0.50, 0.75, 1.0, 1.5 and 2.0) of Murashige and Skoog (MS) medium supplemented with 5 mg l(-1) indole butyric acid and 30 g l(-1) sucrose. Quarter-strength MS medium was proven suitable for the production of both root biomass and secondary metabolites [anthraquinone (AQ), phenolics and flavonoids]. With the increasing salt strength, root growth and AQ accumulation decreased significantly. Higher (1.5 and 2 MS) salt strength provoked osmotic stress resulted in more than twofold free proline accumulation than lower salt strength treated roots and induced free radical scavenging activity. Phenylalanine ammonia lyase activity showed a positive correlation in relation to salt strength that leads to an increase in phenol biosynthesis in expense of AQ formation. The elevated catalase (CAT), guaiacol peroxidase (G-POD) and superoxide dismutase activities and decreased ascorbate peroxidase (APX) activities were observed in roots treated with 2.0 MS. On the other hand, APX activity was strongly activated along with considerable increase in CAT activity at 0.25 MS treated culture. However, the joint functions of CAT, G-POD and APX at 0.25 MS treated cultures were efficient to eliminate the potential danger of hydrogen peroxide (H(2)O(2)) as evidenced from low H(2)O(2) accumulation and low level of lipid peroxidation.

Focussed beam reflectance measurement (FBRM) monitoring of particle size and morphology in suspension cultures of Morinda citrifolia and Centaurea calcitrapa.

Laser light scattering technology, as applied in the Lasentec focussed beam reflectance measurement (FBRM) system, was used to characterise two morphologically dissimilar plant cell suspension cultures, Morinda citrifolia and Centaurea calcitrapa. Shake-flask suspensions were analysed in terms of biomass concentration and aggregate size/shape over the course of typical batch growth cycles. For the heavily aggregated C. calcitrapa, biomass levels [from 10-160 g fresh weight (fw) l(-1))] were linearly correlated with FBRM counts. For M. citrifolia, which grows in unbranched chains of 2-10 elongated cells, linear correlation of biomass concentration with FBRM counts was applicable in the range 0-100 g fw l(-1); at higher levels (100-300 g fw l(-1)), biomass was non-linearly correlated with FBRM counts and length-weighted average FBRM chord length. For both cell systems, particle morphology (size/shape) was quantified using semi-automated digital image analysis. The average aggregate equivalent diameter (C. calcitrapa) and average chain length (M. citrifolia), determined using image analysis, closely tracked the FBRM average chord length. The data clearly demonstrate the potential for applying the FBRM technique for rapid characterisation of plant cell suspension cultures.

[Diagnosis and integrative evaluation on soil fertility of three Chinese medicinal materials in GAP plots].

The fertility of soil was diagnosed and integratively evaluated by modified Nemoro Index with eight indexes, i.e. pH, organic matter, total N, P, K and available N, P, K on three national Chinese medicinal materials GAP (Good Agricultural Practice) experimental plots of Pogostemon cablin, Citrus medica and Morinda officinalis in Guangdong. The results showed that the pH, organic matter and other major nutrient contents were low and imbalance. There are lots of nutritive disturbance factors. The level of soil integrative fertility in any of the plots was poor.

[Effects of various organic fertilizers on growth and mineral nutrient absorption of Morindar officinalis].

OBJECTIVE: To study the effects of various organic fertilizers on the growth and the mineral nutrient absorption of Morindar officinalis. METHOD: The growing character and the mineral nutrition contents were determined with conventional assay methods. RESULT: The influences of four kinds of fertilizers on the bionomics and nutrient absorption of M. officinalis successively were: biotic organic fertilizer A > biotic organic fertilizer C > biotic organic fertilizer B > farm manure. CONCLUSION: Different kinds of organic fertilizers could facilitate the growth and the mineral nutrient absorption of M. officinalis. Increasing the fertilizer amount of germ manure could promote the absorption of mineral nutrients and the accumulation of dry matters in plants.

[In vitro culture and the Agrobacterium-mediated genetic transformation of Morinda officinalis].

OBJECTIVE: To establish an effective system for the Agrobacterium-mediated genetic transformation of M. officinalis, for laying a foundation for the improvement of breeds and introduction of foreign objective genes. METHOD: The explants used for culture were the nodular stem segments from M. officinalis. Agrobacterium tumefaciens strain was EHA101, containing vector plasmid pGA482GG. The GUS gene and NPT II gene were introduced into the plasmid. RESULT: MT basal medium with BA 1 mg.L-1 was effective to inducing the direct shoot formation, and the frequency of shoot formation was 97.8%. As BA concentrations increased, the ability of shoot formation decreased. The explants oriented with their apical ends protruding from the medium produced more shoots than when they were placed with their basal end upright or were placed horizontally. The optimal rooting medium for regenerating shoots was MT basal medium supplemented with 0.2 to 0.5 mg.L-1 NAA, and a root induction rate over 80.0% was observed. The selection pressure for kanamycin was 50 mg.L-1. Cefotaxime was used as antibiotics, and the concentration was 300 mg.L-1. After 1.5 months, 14.8% resistant shoots were emerged from the explants. Histochemical GUS assay showed that 22.2% of the resistant plants were GUS-positive. CONCLUSION: Plant regeneration system and Agrobacterium-mediated genetic transformation have been established for M. officinalis in vitro.