scholarly journals A biochemical and proteomic approach to the analysis of tomato mutant fruit growth

2021 ◽  
Vol 45 (1) ◽  
pp. 71-85
Author(s):  
Milena Marjanovic ◽  
Zorica Jovanovic ◽  
Biljana Vucelic-Radovic ◽  
Sladjana Savic ◽  
Ivana Petrovic ◽  
...  

To assess the effects of ABA deficiency on tomato fruit growth, the ABA mutant flacca was grown in an optimal soil water regime and various analyzes were performed, including morphological (fruit number, diameter and fruit biomass), physiological (duration of growth and fruit growth rate), biochemical (ABA accumulation, enzyme cell wall peroxidase activity) as well as proteomics. The fruit growth analysis showed that the slower fruit growth rate and development resulted in smaller flacca fruits in comparison to the wild-type fruits. The comparison of the temporal dynamics of cell wall peroxidase activity and ABA content in our experiment indicated an opposite relationship during fruit development. Proteomic analysis and the down-regulation of most proteins from carbon and amino acid metabolism, the translation and processing of proteins, energy metabolism and cell wall-related metabolism in the flacca fruits compared to the wild type, indicated reduced metabolic flux which reflected a slower fruit growth and development and reduced fruit size in the ABA mutant. These findings also indicated that ABA limited carbon sources, which could be responsible for the reduced fruit growth and size of ABA-deficient tomato fruits. The up-regulation of sulfur and oxygen-evolving enhancer proteins in the flacca fruits implicated the maintenance of photosynthesis in the late expansion phase, which slows down transition to the ripening stage. The majority of antioxidative and stress defence proteins were down-regulated in the flacca fruits, which could be related to the role of ABA in the activity of different antioxidative enzymes as well as in regulating cell wall expansion and the cessation of fruit growth.

Author(s):  
Dien Thi Kieu Pham ◽  
Kiet Thuong Do ◽  
Sanh Du Nguyen

The cherry tomato fruit size depends on the growth of the pericarp which is parenchymal cells. The blue light stimulates the expansion of cotyledon cells, hypocotyl cells and leaf cells. In this study, the cherry tomato fruit was used as a material to investigate the effects of the blue light on the pericarp cells growth in fruit growth stage and lycopene accumulation in fruit growth and ripening stage. After 7 days of the blue light (440, 450 or 460 nm) treatment, pericarp cells growth and physiological, biochemical changes of the pericarp cells of 7-day-old fruit pericarp piece in vitro were analyzed. The lycopene content and some organic compound contents of 42-day-old postharvest fruits treated by the blue light similarly in 7 days and 7, 21-day-old fruit wrapped with blue filter (440-510 nm filtered) in 7 days were measured. The results showed that the 450 nm wavelength blue light the increased pericarp thickness of 7-day-old fruits through the increasement of the pericarp cell diameter. The 460 nm wavelength blue light the increased lycopene content of 42-day-old postharvest fruits. The blue filter treatment increased the sugar total content of 7- day-old fruits and increased the lycopene content of 21-day-old fruits.


1967 ◽  
Vol 18 (1) ◽  
pp. 95 ◽  
Author(s):  
DI Jackson ◽  
BG Coombe

The effect of temperature and gibberellic acid (GA3) applications on apricot fruit have been determined by measurements of fruit size and shape, mesocarp cell number, size, and shape, and endogenous gibberellin. Application of heat during the first 10 nights after anthesis increased the initial growth rate of fruit and of cells in the mesocarp and produced more rapid cell division in this tissue. It did not affect final fruit size or the number and diameter of cells in the mesocarp. Higher temperatures did, however, hasten maturity of fruit. GA3 perfused into branches before anthesis produced an increased drop of flower buds and fruit, raised the ratio of flower buds to leaf buds initiated that season, and resulted in elongated pedicels. Initially, fruit growth rate was increased by GA3, but subsequently it was depressed and final size was below normal. These effects on fruit size were mainly due to effects on the rate of cell division. Some differences were noted in the dimensions of cells but final radial cell diameter did not differ from untreated fruit. GA3-treated fruit ripened sooner than controls. Neither heating nor GA3 treatments affected the level of endogenous gibberellin-like substances in the fruit or their RF on paper chromatograms. There were no significant interactions between temperature and gibberellin in any parameter of apricot fruit growth.


2021 ◽  
Author(s):  
Shikha Jindal ◽  
Mahesh S. Iyer ◽  
Poonam Jyoti ◽  
Shyam Kumar Masakapalli ◽  
K V Venkatesh

Global regulatory transcription factors play a significant role in controlling microbial metabolism under genetic and environmental perturbations. A systems-level effect of carbon sources such as acetate on microbial metabolism under disrupted global regulators has not been well established. Acetate is one of the substrates available in a range of nutrient niches such as the mammalian gut and high-fat diet. Therefore, investigating the study on acetate metabolism is highly significant. It is well known that the global regulators arcA and fis regulate acetate uptake genes in E. coli under glucose condition. In this study, we deciphered the growth and flux distribution of E.coli transcription regulatory knockout mutants ΔarcA, Δfis and double deletion mutant, ΔarcAfis under acetate using 13C-Metabolic Flux Analysis which has not been investigated before. We observed that the mutants exhibited an expeditious growth rate (~1.2-1.6 fold) with a proportionate increase in acetate uptake rates compared to the wild-type. 13C-MFA displayed the distinct metabolic reprogramming of intracellular fluxes, which conferred an advantage of faster growth with better carbon usage in all the mutants. Under acetate metabolism, the mutants exhibited higher fluxes in the TCA cycle (~18-90%) and lower gluconeogenesis flux (~15-35%) with the proportional increase in growth rate. This study reveals a novel insight by stating the sub-optimality of the wild-type strain grown under acetate substrate aerobically. These mutant strains efficiently oxidize acetate to acetyl-CoA and therefore are potential candidates that can serve as a precursor for the biosynthesis of isoprenoids, biofuels, vitamins and various pharmaceutical products.


2012 ◽  
Vol 137 (5) ◽  
pp. 294-301 ◽  
Author(s):  
Julienne Fanwoua ◽  
Pieter de Visser ◽  
Ep Heuvelink ◽  
Gerco Angenent ◽  
Xinyou Yin ◽  
...  

To improve our understanding of fruit growth responses to temperature, it is important to analyze temperature effects on underlying fruit cellular processes. This study aimed at analyzing the response of tomato (Solanum lycopersicum) fruit size to heating as affected by changes in cell number and cell expansion in different directions. Individual trusses were enclosed into cuvettes and heating was applied either only during the first 7 days after anthesis (DAA), from 7 DAA until fruit maturity (breaker stage), or both. Fruit size and histological characteristics in the pericarp were measured. Heating fruit shortened fruit growth period and reduced final fruit size. Reduction in final fruit size of early-heated fruit was mainly associated with reduction in final pericarp cell volume. Early heating increased the number of cell layers in the pericarp but did not affect the total number of pericarp cells. These results indicate that in the tomato pericarp, periclinal cell divisions respond differently to temperature than anticlinal or randomly oriented cell divisions. Late heating only decreased pericarp thickness significantly. Continuously heating fruit reduced anticlinal cell expansion (direction perpendicular to fruit skin) more than periclinal cell expansion (direction parallel to fruit skin). This study emphasizes the need to measure cell expansion in more than one dimension in histological studies of fruit.


mBio ◽  
2016 ◽  
Vol 7 (4) ◽  
Author(s):  
Jilong Wang ◽  
Dalai Yan ◽  
Ray Dixon ◽  
Yi-Ping Wang

ABSTRACT A fundamental question in microbial physiology concerns why organisms prefer certain nutrients to others. For example, among different nitrogen sources, ammonium is the preferred nitrogen source, supporting fast growth, whereas alternative nitrogen sources, such as certain amino acids, are considered to be poor nitrogen sources, supporting much slower exponential growth. However, the physiological/regulatory logic behind such nitrogen dietary choices remains elusive. In this study, by engineering Escherichia coli , we switched the dietary preferences toward amino acids, with growth rates equivalent to that of the wild-type strain grown on ammonia. However, when the engineered strain was cultured together with wild-type E. coli , this growth advantage was diminished as a consequence of ammonium leakage from the transport-and-catabolism (TC)-enhanced (TCE) cells, which are preferentially utilized by wild-type bacteria. Our results reveal that the nitrogen regulatory (Ntr) system fine tunes the expression of amino acid transport and catabolism components to match the flux through the ammonia assimilation pathway such that essential nutrients are retained, but, as a consequence, the fast growth rate on amino acids is sacrificed. IMPORTANCE Bacteria exhibit different growth rates under various nutrient conditions. These environmentally related behaviors reflect the coordination between metabolism and the underlying regulatory networks. In the present study, we investigated the intertwined nitrogen metabolic and nitrogen regulatory systems to understand the growth differences between rich and poor nitrogen sources. Although maximal growth rate is considered to be evolutionarily advantageous for bacteria (as remarked by François Jacob, who said that the “dream” of every cell is to become two cells), we showed that negative-feedback loops in the regulatory system inhibit growth rates on amino acids. We demonstrated that in the absence of regulatory feedback, amino acids are capable of supporting fast growth rates, but this results in ammonia leaking out from cells as “waste,” benefiting the growth of competitors. These findings provide important insights into the regulatory logic that controls metabolic flux and ensures nutrient containment but consequently sacrifices growth rate.


2005 ◽  
Vol 4 (1) ◽  
pp. 111-120 ◽  
Author(s):  
Ellen M. Kellner ◽  
Kris I. Orsborn ◽  
Erin M. Siegel ◽  
M. Alejandra Mandel ◽  
Marc J. Orbach ◽  
...  

ABSTRACT 1,3-β-Glucan synthase is responsible for the synthesis of β-glucan, an essential cell wall structural component in most fungi. We sought to determine whether Coccidioides posadasii possesses genes homologous to known fungal FKS genes that encode the catalytic subunit of 1,3-β-glucan synthase. A single gene, designated FKS1, was identified, and examination of its predicted protein product showed a high degree of conservation with Fks proteins from other filamentous fungi. FKS1 is expressed at similar levels in mycelia and early spherulating cultures, and expression decreases as the spherules mature. We used Agrobacterium-mediated transformation to create strains that harbor ΔFKS1::hygB, a null allele of FKS1, and hypothesize that Fks1p function is essential, due to our inability to purify this allele away from a complementing wild-type FKS1 allele in a heterokaryotic strain. The heterokaryon appears normal with respect to growth rate and arthroconidium production; however, microscopic examination of strains with ΔFKS1::hygB alleles revealed abnormal swelling of hyphal elements.


2017 ◽  
Vol 83 (17) ◽  
Author(s):  
Arpan De ◽  
Sumei Liao ◽  
Jacob P. Bitoun ◽  
Randy Roth ◽  
Wandy L. Beatty ◽  
...  

ABSTRACTStreptococcus mutansis known to possess rhamnose-glucose polysaccharide (RGP), a major cell wall antigen.S. mutansstrains deficient inrgpG, encoding the first enzyme of the RGP biosynthesis pathway, were constructed by allelic exchange. ThergpGdeficiency had no effect on growth rate but caused major defects in cell division and altered cell morphology. Unlike the coccoid wild type, thergpGmutant existed primarily in chains of swollen, “squarish” dividing cells. Deficiency ofrgpGalso causes significant reduction in biofilm formation (P< 0.01). Double and triple mutants with deficiency inbrpAand/orpsr, genes coding for the LytR-CpsA-Psr family proteins BrpA and Psr, which were previously shown to play important roles in cell envelope biogenesis, were constructed using thergpGmutant. There were no major differences in growth rates between the wild-type strain and thergpG brpAandrgpG psrdouble mutants, but the growth rate of thergpG brpA psrtriple mutant was reduced drastically (P< 0.001). Under transmission electron microscopy, both double mutants resembled thergpGmutant, while the triple mutant existed as giant cells with multiple asymmetric septa. When analyzed by immunoblotting, thergpGmutant displayed major reductions in cell wall antigens compared to the wild type, while little or no signal was detected with the double and triple mutants and thebrpAandpsrsingle mutants. These results suggest that RgpG inS. mutansplays a critical role in cell division and biofilm formation and that BrpA and Psr may be responsible for attachment of cell wall antigens to the cell envelope.IMPORTANCEStreptococcus mutans, a major etiological agent of human dental caries, produces rhamnose-glucose polysaccharide (RGP) as the major cell wall antigen. This study provides direct evidence that deficiency of RgpG, the first enzyme of the RGP biosynthesis pathway, caused major defects in cell division and morphology and reduced biofilm formation byS. mutans, indicative of a significant role of RGP in cell division and biofilm formation inS. mutans. These results are novel not only inS. mutans, but also other streptococci that produce RGP. This study also shows that the LytR-CpsA-Psr family proteins BrpA and Psr inS. mutansare involved in attachment of RGP and probably other cell wall glycopolymers to the peptidoglycan. In addition, the results also suggest that BrpA and Psr may play a direct role in cell division and biofilm formation inS. mutans. This study reveals new potential targets to develop anticaries therapeutics.


2000 ◽  
Vol 122 (2) ◽  
pp. 527-534 ◽  
Author(s):  
Carmen Catalá ◽  
Jocelyn K.C. Rose ◽  
Alan B. Bennett

HortScience ◽  
2012 ◽  
Vol 47 (11) ◽  
pp. 1580-1585 ◽  
Author(s):  
Todd C. Einhorn ◽  
Janet Turner ◽  
Debra Laraway

Reflective fabric was installed before bloom in 2009 and 2010 in alleyways of a mature, low-density ‘Anjou’ pear orchard (269 trees/ha). Four treatments were applied to study intracanopy light environments on fruit growth rate and size, cropload, yield, and fruit quality: 1) no fabric (NF); 2) partial-season fabric applied before full bloom (FB) and removed 75 days after full bloom (dafb) (PSF); 3) full-season fabric applied before FB and removed at harvest (FSF); and 4) shadecloth (60%) applied 60 dafb through harvest (SC). PSF and FSF improved yield by 12% and 18%, respectively, over the two-year period relative to NF. The high yields of fabric treatments were attributed to fruit number in the lower (less than 2.4 m) interior, mid-, and exterior zones of the canopy. Photosynthetic active radiation (PAR) was increased by fabric 28%, 95%, and 30% in the lower exterior, mid-, and interior canopy, respectively. Photosynthesis:light response curves indicated improved carbon assimilation of pear leaves developing in the elevated PAR environment of the lower canopy. Fruit growth rate and final size were unaffected by fabric treatments. FSF fruit size was similar to NF despite higher fruit density. Compared with NF, FSF had a small, non-significant effect on fruit maturity (increased softening) at harvest. Yield and fruit size of SC fruit were significantly reduced. The number of fruit in SC trees did not differ from NF in 2009, but the effect of shade reduced fruit number in 2010. Fabric did not affect fruit quality attributes after three and six months of regular atmosphere cold storage. Pears from SC trees did not attain ripening capacity after three months of cold storage and a 7-day ripening period and had lower sugar content compared with other treatments. The cumulative yield advantages associated with FSF support its use in mature pear orchards.


2012 ◽  
Vol 39 (No. 1) ◽  
pp. 26-32 ◽  
Author(s):  
M.M. Hossain ◽  
H. Nonami

The effect of salt stress on physiological response of hydroponically grown tomato fruit was investigated. Fruit growth rate, water status, cuticle permeability and induction of blossom-end rot (BER) of tomato fruit were considered for this study. Salt stress was applied by using Ca salt treatment and it plays an important role on all parameters studied in this experiment. Fruit growth rate, predawn water potential, osmotic potential and cuticle permeability were significantly lower in treated plants than in control plants. On the other hand, tissue turgor of control and treated fruit showed almost similar values 12 days after flowering (DAF). This result indicated that turgor was osmotically regulated in fruit under stress condition. Fruit growth rate was found to decline from 12 DAF and eventually ceased when BER externally appeared on fruit surface at the age of 19 DAF in this experiment. The reduction of growth rate coincided with the reduction of water potential in fruit tissue due to salt stress. Although BER externally appeared at 19 DAF anatomical investigation showed that intercellular air space becomes discoloured at least one week before external symptoms appeared on fruit tip. Different levels of cuticular permeability indicated that the deposition of cuticular wax on fruit surface was enhanced by the salt stress condition in tomato fruit. Since, BER was found to appear on fruit tip under high calcium concentration in solution it can be concluded that calcium deficiency was not the only the cause of BER in tomato, rather salt stress might alter metabolic activity in developing tomato fruit.


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