Bacterial soft rot of tomato in plastic greenhouses in Crete

During recent years a new disease has been noticed on tomatoes grown in Polythene greenhouses in Crete. Early symptoms are yellowing of the lower leaves, and a yellow brown discoloration of the pith and stem xylem. As leaves wilt and die there is progressive yellowing towards the top of the plants. A progressive disintegration of the cortical tissues follows which results in a soft rot and a longitudinal splitting of the stem running mainly upwards. Soft rot of the fruits rarely appears. Severely infected plants may wilt and die, but other less affected plants often survive and yield normally. Very vigorous plants grown under humid conditions are more susceptible. Often more than 20% of the plants are infected. Isolations were made from stem (xylem, cortex and pith), from leaf xylem and from fruits of infected tomato plants collected throughout the island from 1979 to 1985. Bacteria of the genus Erwinia and Pseudomonas were consistently isolated. On the basis of physiological and biochemical characters of 49 representative pathogenic isolates, 22 were identified as Erwinia carotovora subsp. carotovora, 10 as Erwinia carotovora subsp. atroseptica, four as Pseudomonas viridiflava and 13 as Pseudomonas fluorescens biotype I. All disease symptoms were reproduced when artificial inoculations were made with the above isolates in the laboratory (20°C and 100% r.h.) on 3–4 week tomato plants and in a commercial greenhouse on 4–5 months tomato plants. Bacteria used for inoculations were reisolated. Results indicated that the disease symptoms as described may be caused by four different bacteria species.     

The epidemiology of tomato brown root rot

In the absence of nematodes, three different symptoms of disease, parts of the brown root rot complex (BRR), occurred on tomato roots surviving in soils infested with GSF (= grey sterile fungus) and Colletotrichum atramentarium (Berk. & Br.) Taubenh. In heavily infested soils brown lesions occurred throughout cropping, appearing within a week of planting. Corkiness and black dot, caused by GSF and C. atramentarium respectively, rarely occurred until the third month after planting but towards the end of the season the incidence of black dot sometimes suddenly increased greatly. Observations of crops growing in plots treated with different soil partial sterilants suggested that GSF was more damaging than C. atramentarium. Yield was not related to the incidence of black dot but was inversely proportional to the occurrences of brown lesions and corkiness. The relation with brown lesions was significant within 8 weeks of planting, when most brown lesions gave cultures of GSF, but later more of these lesions gave cultures of C. atramentarium than of GSF. Pathogenicity tests with pure cultures of GSF and C. atramentarium were done on agar media and by artificially infesting partially sterilized soils. Roots of undamaged seedlings on agar media developed 10 mm. brown lesions within 2 weeks of inoculating 10-day-old tomatoes with most GSF cultures isolated from: (1) rotted roots of Lycopersicon esculentum, Solanum capsicastrum, Capsicum annuum var. longum and C. frutescens; (2) browned zones of Lycopersicon hirsutum roots; and (3) apparently healthy roots of Cucumis sativus. After inoculation with C. atramentarium, small (c. 2 mm.) pink lesions developed, whereas none formed using Pyrenochaeta spp. In soil tests the greater root damage done by GSF, including root loss, was reflected in decreased aerial growth and smaller fruit yields; C. atramentarium affected neither. In the second year of soil infestation GSF decreased yields during 6 weeks of picking from 1.96 kg. in the uninoculated controls to 1.02 kg./plant. The pattern of damage done by GSF changed as plants aged. In soil, brown lesions occurred within a few days of planting but corkiness did not appear for 2–3 months, when stem lesions and leaf yellowing often developed simultaneously. A 50% root loss after 21 weeks did not affect fruit yields whereas a 40% loss within 11 weeks of planting was reflected by a 45% yield decrease.     

Phytophthora cryptogea root rot of tomato in rockwool nutrient culture:

The effect of root temperature on growth and yield of rockwool-grown tomato plants infected with Phytophthora cryptogea was investigated. Measurements of shoot and root growth were taken at high (25^oC) and low (15^oC) root temperatures during the generative phase of growth. The growth of roots of healthy and P. cryptogea-infected tomato plants in rockwool blocks was higher in plants grown with roots at 25^oC than at 15^oC after 60 days and a similar effect was found in slabs after 98 days. Under sub-optimal conditions for growth the disease became severe when root temperatures were low. Growth of roots was greatest when roots were maintained at a high temperature in combination with an ambient air temperature of c. 15^oC and the response was greater in cv. Counter than cvs Calypso and Marathon. Water-soluble carbohydrates of roots were higher in those produced in blocks than slabs and were reduced by infection compared to healthy plants with roots at 15^oC and 25^oC. Reduced transpiration rates were found 17 days after inoculation in symptomless plants grown at a root temperature of 25^oC. Infection, regardless of the temperature of the roots or cultivar, led to reduced stem growth. The plants grown at 25^oC were taller than those with a root temperature of 15^oC. After 9 wk of harvest, the cumulative fruit yields in infected cvs Counter and Calypso grown at 25^oC were comparable to that in healthy plants grown at either temperature and cumulative fruit numbers followed a similar pattern. High root temperatures led to delayed fruit ripening between weeks 3–10 and a larger number of unripe fruit. The weight of unripe fruit from infected plants grown at 25^oC at the terminal harvest was higher than from healthy plants with roots maintained at 15^oC.     

Build-up of tomato brown root rot caused by Pyrenochaeta lycopersici Schneider and Gerlach

The incidence in unheated glasshouses of tomato brown root rot (BRR) was recorded on five successive crops starting with the first ever grown on the experimental site. In the protected environment the changes followed a pattern that was defined mathematically, the regression linking angular or logit transforms of per cent BRR, observed at different intervals after planting, accounting for c. 72% of the variation. Rates of build-up, seemingly related to initial amounts of inocula, increased in successive seasons, there being more BRR in the fifth season than in the first. Four per cent BRR recorded 5 weeks after planting increased threefold during the next 3 months to 12 %, but in later years, when amounts of early-season infection were doubled to 8%, the incidence of root rot subsequently increased five times. Tomato roots colonized by Pyrenochaeta lycopersici either brown and rot or become corky. The proportion of end-of-season BRR attributable to corkiness increased from the first to fifth seasons. The monthly increments of fruit yield were inversely proportional to the incidence of BRR.     

Phytophthora cryptogea root rot of tomato in rockwool nutrient culture. I. Analysis of root infection

The spatial and temporal colonisation of tomato plants by Phytophthora cryptogea was studied in rockwool nutrient culture. Root growth and the distribution and progress of infection were measured on dissected root fragments obtained from a detailed destructive sampling of the substrate. Dissected fragments of root or roots and fibres were plated on BNPRA agar, a selective Phytophthora medium. Roots colonised all parts of the rockwool substrate 60 days after planting with the exception of surface marginal and central areas of the slab which had a lower solution content. Most root biomass occurred in and immediately beneath the original growing block. The distribution of P. cryptogea closely followed the pattern of root colonisation. An alternative, novel method for root analysis involved the dissolution of the mineral fibre and its formate bonding resin by digestion in 1 m H₃PO₄ for 45 min. Comparative recoveries of P. cryptogea from plated fragments of dissected root and fibre or comminuted samples of acid-released or dissected root showed that the acid treatment initially reduced the number of Phytophthora colonies in block and slab roots by 67% and 61% respectively. After 28 days, colony recovery from acid-released roots in the rockwool slab increased and was between 4% and 13% lower than from other plating methods. Since 1 m H₃PO₄ was lethal to zoospores and surface sporangia, the colonies recovered were interpreted as originating exclusively from root lesions. Root fresh weight of healthy and inoculated plants declined during the initial period of fruit formation. P. cryptogea infection led to a progressive reduction in root weight in the growing block and main slab and 28 days after inoculation, was approximately 50% of the controls. The acid digestion of rockwool fibre is proposed as a new approach to the problem of root and pathogen analysis in this substrate.     

A formulation of neem cake seeded with Bacillus sp. provides control over tomato Fusarium crown and root rot

Fusarium crown and root rot (FCRR) caused by the fungus Fusarium oxysporum f. sp. radicis-lycopersici is a damaging soil-borne disease of tomato. A plant growth rhizobacterium, Bacillus sp. strain HN09 isolated from neem tree rhizosphere soil, was shown to inhibit the growth, germination and development of normal morphology of the FCRR pathogen. A substantial level of disease control was achieved in greenhouse trials by soil supplementation with a preparation of neem cake seeded with HN09. Dry sterilisation of neem cake before fermentation gained comparable disease control effect as those in the unsterilised treatment, whereas moist sterilisation treatment decreased the effect significantly. This bioformulation also led to significantly raised activities in tomato genes encoding pathogenesis-related proteins, hence providing an effective alternative for the control of FCRR, reducing the need for chemical fungicide and fertilisers that impact the environment.     

Effects of cultural treatments on the incidence of, and damage done by, tomato brown root rot

In soils naturally infested with Pyrenochaeta lycopersici, which usually occurs as a grey sterile fungus (GSF), symptoms of brown root rot (BRR) developed sooner and more extensively in the second year of cropping than in the first. The amount of BRR attributable to corkiness increased as plants aged but, at comparable stages of cropping, decreased in the second and third seasons, an effect associated inversely with the severity of early GSF attack. The larger amount of corkiness, 33 compared with 18 %, on two batches of plants in 1965, each with 53 % end-of-season BRR, was also attributed to a less severe early GSF attack on the former than on the latter, growing in soils unsteamed for 1 and 3 years respectively. The incidence of BRR decreased with increasing depth in infested soils but increased on plants grown in plots with partially sterilized topsoil. Partially sterilizing soils at the G.C.R.I. and Fairfield E.H.S. decreased the incidence of BRR and increased crop weights from about the second month of picking, but fruit quality was poorer. Seasonal yields from plants in untreated soil progressively decreased relatively to those from repeatedly steamed plots, from 93 to 65 % in successive years at Fairfield and from 65 to 56 and 43 % at the G.C.R.I. Steaming done in 1963 and 1964 temporarily retarded GSF attack in 1965 with corresponding yield increases. Increasing the amounts of sterilized propagating soil surrounding roots at planting from 0·4 to 1·01 per plant increased yields by c. 0·4 kg/plant, this being a relatively large increase for plants in infested soil, where this treatment significantly delayed the early incidence of BRR near stem bases. Grafting commercially acceptable scions to rootstocks that tolerated colonization by GSF (‘resistant’ rootstocks), temporarily checked growth, delayed picking and decreased fruit quality. Usually grafted plants, irrespective of soil treatment, yielded at least as much fruit as ungrafted plants in steamed soil. In one of five comparisons, soil steaming increased yields of grafted plants. When testing the effects of previous cropping it seemed that populations of GSF increased similarly in soils planted with grafted and ungrafted plants. In addition to GSF attack, roots at Fairfield E.H.S. were often colonized by Colletotrichum coccodes, microsclerotia (= black dots) being more numerous as plants aged. Although significantly more black dot developed on GSF-resistant rootstocks grown in untreated soil than on those grown in steamed soil, the differences were not associated with effects on yield. C. coccodes colonized GSF-susceptible and -resistant roots equally.     

Biological control of strawberry crown rot,root rot and grey mould by the beneficial fungus Aureobasidium pullulans

Utilization of biocontrol agents is a sustainable approach to reduce plant diseases caused by fungal pathogens. In the present study, we tested the effect of the candidate biocontrol fungus Aureobasidium pullulans (De Bary) G. Armaud on strawberry under in vitro and in vivo conditions to control crown rot, root rot and grey mould caused by Phytophthora cactorum (Lebert and Cohn) and Botrytis cinerea Pers, respectively. A dual plate confrontation assay showed that mycelial growth of P. cactorum and B. cinerea was reduced by 33–48% when challenged by A. pullulans as compared with control treatments. Likewise, detached leaf and fruit assays showed that A. pullulans significantly reduced necrotic lesion size on leaves and disease severity on fruits caused by P. cactorum and B. cinerea. In addition, greenhouse experiments with whole plants revealed enhanced biocontrol efficacy against root rot and grey mould when treated with A. pullulans either in combination with the pathogen or pre-treated with A. pullulans followed by inoculation of the pathogens. Our results demonstrate that A. pullulans is an effective biocontrol agent to control strawberry diseases caused by fungal pathogens and can be an effective alternative to chemical-based fungicides.

     

Fungicidal drenches for control of root rot in subterranean clover

Fungicide drenches of benomyl, metalaxyl, iprodione, propamocarb, or thiram were applied to intact soil cores taken from known root rot affected fields in Western Australia, to control subterranean clover root rot. Metalaxyl was the most effective in reducing seedling damping-off. The most effective fungicide for reducing the level of rotting of both tap and lateral root systems of survixing plants varied from season to season at one site and varied between different sites in any one season with each fungicide giving a significant reduction in root disease on at least one occasion. Results suggest that different individuals or complexes of root pathogens were operative between seasons in any one site, and between sites for any one season. In some instances it appears that different individual root pathogens or pathogen complexes were operative on tap roots compared to lateral roots.     

Rhizospheric bacteria as potential biocontrol agents against Fusarium wilt and crown and root rot diseases in tomato

The discovery of novel biocontrol agents requires the continuous scrutiny of native microorganisms to ensure that they will be useful on a regional scale. The goal of the present work was to discover novel antagonistic bacteria against Fusarium oxysporum ff. spp. lycopersici race 3 (Fol R3) and radicis-lycopersici (Forl) causing Fusarium wilt disease and Fusarium crown and root rot of tomatoes, respectively. High-throughput liquid antagonism screening of 1,875 rhizospheric bacterial strains followed by dual confrontation assays in 96-well plates was used to select bacteria exhibiting > 50% fungal growth inhibition. In a second dual confrontation assay in 10-cm Petri dishes, bacteria showing > 20% Fol R3 or Forl growth inhibition were further screened using a blood hemolysis test. After discarding β-hemolytic bacteria, a seedling antagonistic assay was performed to select five potential antagonists. A phylogenetic analysis of 16S rRNA identified one strain as Acinetobacter calcoaceticus (AcDB3) and four strains as members of the genus Bacillus (B. amyloliquefaciens BaMA26, Bacillus siamensis BsiDA2, B. subtilis BsTA16 and B. thuringiensis BtMB9). Greenhouse assays demonstrated that BsTA16 and AcDB3 were the most promising antagonists against Fol R3 and Forl, respectively. Pathogen biocontrol and growth promotion mechanisms used by these bacteria include the production of siderophores, biofilm, proteases, endoglucanases and indole acetic acid, and phosphate solubilization. These five bacteria exerted differential responses on pathogen control depending on the tomato hybrid, and on the growth stage of tomatoes. We report for the first time the use of an Acinetobacter calcoaceticus isolate (AcDB3) to control Forl in tomato under greenhouse conditions.     

Biological control of the root rot and wilt diseases ofLens culinaris medic

Summary Biological control visualizes a control of disease with the help of living organisms. In case of root disease, organisms occur in soil which suppress the activity of disease fungi. In the present investigation antagonists have been isolated and used in the control of root rot and wilt disease ofLens culinaris caused byS. rolfsii andF. oxysporum respectively.Isolation of antagonistic micro-organisms were done from the rhizosphere soil of Bombay 20 and local varieties and from field soil. Antagonists were screened against certain test organisms and pathogens. The selected antagonists were used for the control of the diseases of lentil crop by inoculating the antagonists in the soil and thereby to see the effect of decreased plant disease in sterilized and unsterilized soil.Amendments in the form of different organic materials were also given to control the diseases in natural soil.The most effective results were obtained withT. viride, Streptomyces gougeroti Streptomyces species 12 and 13, bacterial isolates no. 16 and 18 in case ofF. oxysporum f.lentis inoculated pots, where there was no disease. In case ofS. rolfsii inoculated pots not a single antagonist was able to eradicate the disease completely.Portion of the Ph. D. thesis accepted by the University of Saugor, Sagar, India.     

Collimonas fungivorans, an unpredicted in vitro but efficient in vivo biocontrol agent for the suppression of tomato foot and root rot

Although bacteria from the genus Collimonas have demonstrated in vitro antifungal activity against many different fungi, they appeared inactive against the plant-pathogenic fungus Fusarium oxysporum f.sp. radicis-lycopersici (Forl), the causal agent of tomato foot and root rot (TFRR). Visualization studies using fluorescently labelled organisms showed that bacterial cells attached extensively to the fungal hyphae under nutrient-poor conditions but not in glucose-rich Armstrong medium. Collimonas fungivorans was shown to be as efficient in colonizing tomato root tips as the excellent colonizer Pseudomonas fluorescens strain WCS365. Furthermore, it appeared to colonize the same sites on the root as did the phytopathogenic fungus. Under greenhouse conditions in potting soil, C. fungivorans performed as well in biocontrol of TFRR as the well-established biocontrol strains P. fluorescens WCS365 and Pseudomonas chlororaphis PCL1391. Moreover, under biocontrol conditions, C. fungivorans did not attach to Forl hyphae colonizing plant roots. Based on these observations, we hypothesize that C. fungivorans mainly controls TFRR through a mechanism of competition for nutrients and niches rather than through its reported mycophagous properties, for which attachment of the bacteria to the fungal hyphae is assumed to be important.     

Phytophthora cryptogea root rot of tomato in rock woo] nutrient culture. II. Effect of root zone temperature on infection, sporulation and symptom development

The effect of root-zone temperature on Phytophthora cryptogea root rot was studied in tomato cv. Counter grown under winter and summer conditions in rockwool culture. A nutrient temperature of 25°C resulted in increased root initiation and growth, higher in winter-grown than in summer-grown plants. Rhizosphere zoospore populations were greatly reduced at 25°C and above. Growth of P. cryptogea in vitro was optimal between 20°C and 25°C and completely suppressed at 30°C. Encystment was enhanced by increased temperatures above 20°C. Zoospore release in vitro occurred in cultures maintained at constant temperatures in the absence of the normal chilling stimulus. Optimal release was at 10°C; no zoospores were released at 30°C. Inoculated, winter-grown tomato plants maintained at 15°C developed acute aerial symptoms and died after 21 days. Comparable plants grown at a root-zone temperature of 25°C remained symptomless for the 3-months duration of the experiment. Summer-grown infected plants at the higher root temperature wilted but did not die. Enhanced temperature was ineffective as a curative treatment in summer-grown plants with established infection. Aerial symptoms of Phytophthora infection are seen as a function of the net amount of available healthy root. With high root zone temperatures this is determined by new root production and decreased inoculum and infection.     

The occurrence of root rot and crown rot of rice in Gilan and Zanjan provinces, Iran

Root rot and crown rot of rice is one of the important fungal diseases of rice in Gilan and Zanjan provinces, Iran. During 1999--2002, samples of plant and soil around the roots of infected rice plants were collected and used to identify the causal agent. Root and crown parts were surface sterilized with sodium hypochlorite and then cultured on PDA (potato dextrose agar), PPA (pepton pentacholoritobenzene agar) and CLA (carnation leaf agar) media. Soil samples prepared in water agar were used to isolate the pathogen. The causal agent was identified as Fusarium moniliforme. Colonies were initially white but turned violet to grey late. Microconidia were arranged in chain and macroconidia were cylindrical and long with 3-5 septa. The disease was severe in Zanjan province particularly along Ghezel Ozan river where the infection ranged from 70-80%. Root and crown rot was more prevalent in areas where Champa and Gerdeh were being cultivated continuously. On the other hand, Sadri cultivars had relatively less infection. Persistent cultivation of rice and seed sowing method intensified disease development and caused significant economic losses.     

Biological control of root rot-root knot disease complex of tomato

Efficacy of Pseudomonas aeruginosa alone or in combination with Paecilomyces lilacinus was evaluated in the control of root-knot nematode and root-infecting fungi under laboratory and field conditions. Ethyl acetate extract (1 mg/ml) of P. lilacinus and P. aeruginosa,respectively, caused 100 and 64% mortality of Meloidogyne javanica larvae after 24 h. Ethyl acetate fractions of biocontrol agents were more effective than hexane extracts in the suppression of M. javanica larvae, indicating that active nematicidal compounds are intermediary in polarity. In field experiments, biocontrol fungus and bacterium significantly suppressed soilborne root-infecting fungi including Macrophomina phaseolina, Fusarium oxysporum, Fusarium solani, Rhizoctonia solani and Meloidogyne javanica, the root-knot nematode. P. lilacinus parasitized eggs and female of M. javanica and this parasitism was not significantly influenced in the presence of P. aeruginosa. P. aeruginosa was reisolated from the inner root tissues of tomato, whereas P. lilacinusdid not colonize tomato roots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Response of faba bean to intercropping,biological and chemical control against broomrape and root rot diseases

Multispecies cropping systems contribute to sustainable agriculture with multiple ecosystem services. Effects of intercropping of various crops with faba beans on growth and yield parameters and disease severity of root rot, damping off and broomrape were investigated. This study was implemented in the laboratory, greenhouse and field to investigate the effect of the intercropping systems (fenugreek + faba bean, lupine + faba bean, garlic + faba bean and sole faba bean). The intercropping systems were combined with the application of arbuscular mycorrhiza fungi (AMF) and yeast as bio-control agents, compared to chemical application of herbicides (Glyphosate) and fungicides (Rizolex-T50), to control rot root diseases and broomrape weeds, Orobanche spp., of faba bean plants in vivo and under the naturally infested field. In vitro, yeast and Rizolex-T50 significantly inhibited mycelial growth of root pathogenic fungi. Intercropping with garlic and/or application of Rizolex-T, significantly decreased the incidence and disease index of root rot and damping-off diseases, meanwhile increased percentage of survival plants. In vivo, intercropping with fenugreek and/or application of Glyphosate, significantly reduced the number/weight of spikes/plot of broomrapes. Intercropping with fenugreek combined with AMF application promoted crop growth and significantly increased yield components. The AMF enhanced seed yield/ha when applied to the intercropping of faba bean + fenugreek and faba bean + garlic, showing the highest seed yield/ha with 3.722 and 3.568 ton/ha, respectively. Intercropping of faba bean with garlic integrated with AMF revealed the highest values of LER, 2.45, and net return, 2341 US$/ha. Our results suggested that using faba bean–garlic intercrop along with AMF inoculation can reduce root rot disease, damping off and broomrapes, as well as enhance the profitability of Egyptian farmer and sustainable production.     

Pythium root rot of crucifers

A destructive root disease of cabbage (Brassica oleracea L. var. capitata) and cauliflower (Brassica oleracea L. var. botrytis) incited by a species of Pythium Pringsheim is described as occurring in Varanasi, U.P. The pathogen was isolated on potato dextrose and corn meal agar. Pathogenicity and host range of the disease were studied. Cultural characters, morphology and developmental stages and life cycle of the fungus indicated its identity with Pythium middletonii Sparrow.     

Aphanomyces root rot of cauliflower

Résumé L'influence du cycloheximide sur la composition chimique des parois de deux levures Rhodotorula après 24 heures et 4 jours de croissance dans deux milieux de culture différents a été étudiée.La composition en glucides neutres ne varie que faiblement en fonction de l'âge des cellules, la détermination du rapport des oses neutres n'indiquant qu'un léger enrichissement en mannose dans les parois des cellules âgées. Chez Rhodotorula glutinis, les teneurs en hexosamines et aminoacides augmentent nettement en fonction de l'âge des cellules, cette variation ne se produit pas chez Rhodotorula rubra. Les teneurs en chitine sont dépendantes de la nature du milieu de culture mais non de l'âge des cellules sauf chez Rhodotorula glutinis.L'influence du cycloheximide sur la composition chimique se traduit par une baisse globale en protéines et par une perturbation des concentrations d'aminoacides.Le rapport molaire des oses neutres indique une baisse systématique du galactose. Les valeurs des hexosamines sont également plus faibles; la diminution de la chitine est surtout très nette lorsque les cellules se développent dans un milieu normalement favorable à son élaboration, cependant il n'apparaît pas de relation directe entre les baisses des teneurs en hexosamines et en chitine.

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Summary The influence of cycloheximide on the chemical composition of the cell walls of two Rhodotorula yeasts, cultivated during 24 and 96 hours in two different media, has been studied.The composition, of the neutral sugars varies only weakly with the age of the cells, the determination of the molar ratio revealing only a little increase of mannose.Hexosamine and aminoacids contents in Rhodotorula glutinis cells distinctly increase in old cells; this variation does not appear in Rhodotorula rubra. The content of chitine depends on the culture conditions but not on the age of cells, except with Rhodotorula glutinis.The influence of cycloheximide on the chemical structure is responsable of a general decrease of protein and a perturbation of the concentration of the different aminoacids. The molar ratio of the neutral sugars indicates a decrease of galactose in the cell walls. The values of hexosamine are also lower; the decrease of chitine is especialy appearant when the cells are cultivated in a medium which normaly favors the synthesis of this polymer, but there is no direct relation between the decrease of hexosamin and chitin.