Transmission of Spiroplasma citri by the aster leafhopper Macrosteles fascifrons (Homoptera: Cicadellidae)

The aster leafhopper (Macrosteles fascifrons), injected with an isolate of Spiroplasma citri obtained from brittle root-diseased horseradish (Armoracia rusticana), transmitted the spiroplasma to horseradish and China aster (Callistephus chinensis.) After feeding on plants infected with S. citri, M. fascifrons transmitted the spiroplasma from aster to aster and horseradish, from yellow rocket (Barbarea vulgaris) to aster, and from turnip (Brassica rapa) to turnip. Symptoms in infected horseradish were chlorosis and stunting of newly formed leaves, discoloration of root phloem, and reduced plant growth typical of brittle root disease. Chlorosis, stunting, and asymmetry of young leaves occurred in affected aster and turnip. Flowers of infected aster were small and pale in colour and occasionally showed other symptoms including asymmetry, petal distortion, or light green petals. Spiroplasmas were isolated from all plants showing symptoms. Transmission rates by M. fascifrons which acquired S. citri by feeding on infected plants were very low, but injected leafhoppers transmitted more frequently. This is the first report of the transmission of S. citri from diseased to healthy plants by M. fascifrons.     

Attachment of the Flavescence doree pathogen (MLO) to leafhopper vectors and other insects

Flavescence doree (FD) is an important yellows disease of grapevine, caused by mycoplasma-like-organism (MLO) and is transmitted in the field by the leafhopper Scaphoideus titanus Ball. It can be transmitted in the laboratory between Vicia faba test plants by the leafhopper, Euscelidius variegatus Kbm. A technique to identify a specific attachment system between the MLO and the leafhopper vectors was developed. In this method, called “Double Dot”, extracts of macerated healthy whole insects or organs applied to a support membrane or cryosections of healthy whole leafhoppers, are incubated with a MLO-enriched extract from FD-infected V. faba or FD-infected E. variegatus. Attached MLO cells were identified by immunolabelling using FD-MLO specific monoclonal antibodies. Attachment of MLO cells was obtained on extracts of healthy S. titanus and E. variegatus and on tissues such as salivary glands, hemolymph and alimentary tract. On cryosections, MLO attachment was obtained on acini IV and V of the salivary glands and on some acini III, on the ventriculus of the alimentary tract, and on the abdomen fat bodies. “Double dot” experiments were done using other insect species, and MLO cells attachment was obtained on most MLO-vector insects but also on insects from a few non-vector species.     

Préparation des Antigenes des Mycoplasmes (MLO) Pathogènes de la Flavescence Dorée, à Partir de Tissus Végétaux

Antigen preparation from plant tissues of pathogenic mycoplasms (MLO) causing flavescence doree disease Extraction and purification of plant yellow's pathogen mycoplasms (MLO) from plant tissues is a difficult problem. It concerns indeed non cultivable, heterogeneous and fragile organisms which are localized in the fibrous and resistant phloem tissue. In a work directed by an infectivity test by leafhopper injection, our laboratory investigated the best host plant and the best extraction method for this type of pathogen. Broad bean, Vicia faba gave better extracts than Vitis vinifera. Stems are better than petioles or lamina. The best results were obtained with the top region of the stem, at the level where symptoms are apparent on young plants. The most efficient mincing method is achieved with razorblades moved alternatively by the rapid vertical movement of an electric knife. The extraction medium already published (Caudwell and Kuszala 1986) has to be modified for plants by various additional components, histidine buffer, antioxidizers (glutathion 0.2 mg per ml) and polymers 0.5 to 1 p 100 PVP or Polyclar, 1–2 p 100 PEG). 1 g of plant infected tissue is minced in 4 ml of medium. The extracts are filtered through a 100 mesh nylon cloth. After this stage the purification method goes parallel to that used for leafhopper extracts (Caudwell and Kuszala 1986). It is possible to obtain 4 × 10⁶ infectious units from 1 g of broad bean stem (for calculation, see C 1986). It is possible to obtain 4 × 10⁶ infectious units from 1 g of broad bean stem (for calculation, see Caudwell 1977). The possibility to extract MLO, either from infectious leafhoppers or from diseased plants enabled cross immunological assays involving antigens from one host and antibodies directed towards the other host antigens. The first step was the successful ISEM visualization of ttie MLOs (Caudwell et al. 1982), the second is the immunoenzymatic MLO-detection (Boudon et al. 1986).     

Leafhopper transmission of Rhynchosia little leaf, a disease associated with mycoplasma-like organisms in Jamaica

Little leaf disease of Rhynchosia minima (RLL) in Jamaica is reported for the first time. The presence of phloem-restricted MLO in diseased but not healthy plants, the remission of symptoms induced in RLL-affected plants with soil drenches of tetracycline, but not penicillin, and the transmission of disease-associated MLO to R. minima test plants, suggests that RLL has an MLO aetiology. RLL is vectored by the cicadellid leafhopper Ollarianus balli, for which R. minima represents the specific field host. Healthy colonies of O. balli produced from eggs oviposited on the RLL-immune weed Asystasia gangetica suggest that RLL is not transovarially transmitted. O. balli acquired the RLL agent after access to infected plants for 5 days (shorter feeds were not tried), and there was a maximum latent period in the leafhopper of 21 days. Of the O. balli collected from heavily-infected field stands of RLL, 35% transmitted the disease, while, of those reared on RLL in captivity for 14–16 days, 56% transmitted. Male and female O. balli transmitted equally efficiently, while nymphs were less frequent vectors. O. balli also infected Cajanus cajan, an important small scale subsistence crop in Jamaica, and Catharanthus roseus. It did not, however, transmit coconut lethal yellowing (CLY) disease to test palms after natural or deliberate acquisition-feeding on RLL, acquisition-feeding on CLY-affected palms, or, after injection with CLY-affected phloem exudate. There was thus no evidence that RLL is related to CLY or that O. balli can act as a vector of CLY.     

Studies of the mycoplasma-like organism (MLO) in spinach leaves affected by the aster yellows disease

Summary Mycoplasma-like organisms (MLO) were found in the phloem of spinach (Spinacia oleracea L.) leaves infected with the agent of aster yellows disease by means of the leafhopperMacrosteles fascifrons Stål. The MLOs occurred mainly in mature sieve elements but were recorded in occasional phloem parenchyma cells as well. The MLO showed the typical features of this organism. The majority were ovoid or spherical, some were irregular in form or elongated. The larger bodies were commonly accompanied by small bodies which appeared to originate from the larger by budding. Profiles suggesting binary fission and filamentous forms containing ovoid condensations of cytoplasm were present. The bounding membrane showed the typical trilaminate structure, and DNA-like fibrils were discernible in those MLOs that had an electron lucent central region. In the denser bodies the fibrils were obscured. The MLO ribosomes were distinctly smaller than those in the host cytoplasm. The MLOs were degenerating in phloem cells that were disorganizing and collapsing in response to the infection. Structures in host cells that may be confused with MLO are described.This work was supported by the National Science Foundation grant GB-35228 to K. E and by Hatch and California Statewide Critical Applied Research Funds to the Departmem of Cell Physiology, University of California, Berkeley, California. The authors thank ProfessorJulius H.Freitag for providing the original strains of the aster yellows agent.     

Use of Polyclonal Antibodies to Identify Mycoplasma-like organisms (MLOs) From the Sudan and from Thailand

Rabbit polyclonal antibodies prepared against faba bean phyllody MLO from the Sudan reacted with its homologous antigen and with extracts of Catharanthus roseus experimentally infected with the same or a related MLO from Crotalaria saltiana showing symptoms of phyllody disease, as well as with extracts of naturally MLO-infected C. saltiana growing in the field in the Sudan. The antibodies also reacted positively with extracts of C. roseus experimentally infected with Crotalaria juncea phyllody MLO and soybean phyllody MLO from Thailand. Polyclonal antibodies prepared against an MLO associated with witches' broom disease in C. juncea reacted positively in ELISA tests with homologous antigen extracts from naturally infected C. juncea as well as with extracts from experimentally infected C. roseus and with extracts prepared from Sesamum indicum plants with phyllody symptoms growing in Thailand. There was no reaction between these antibodies and extracts from C. roseus plants infected with the MLOs associated with C. juncea phyllody or with soybean phyllody. No cross reactions were observed among the antigens and antibodies of the two MLO groups by immunoflorescence, ELISA or western blotting. However, the molecular weight of the principal protein antigen, determined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting was the same for both types of MLO. Serologically-similar MLOs thus occur in the Sudan and in Thailand, where they are associated with phyllody symptoms in C. saltiana and faba bean and with C. juncea and soybean, respectively. A second, serologically distinct MLO group was also found infecting C. juncea and S. indicum in Thailand but MLOs from this group have not yet been identified in crops from the Sudan.     

Epidemiological studies on the stunt disease of highbush blueberry

Population levels of Scaphytopius spp., possible sharpnosed leafhopper vectors of blueberry stunt disease (BBSD), were monitored during 1989,1990 and 1991, using yellow sticky traps and a D-Vac power aspirator. Scaphytopius magdalensis (Prov.), S. frontalis (Van D.) and 5. acutus (Say) had two population peaks, one after the petal fall stage and a larger second peak in late Summer to early Autumn. Healthy cv. Bluecrop highbush blueberry (Vaccinium corymbosum L.) plants were placed under stunt-diseased bushes in the field for 2-wk periods during 1989 and 1990. These plants and some of the leafhoppers trapped during 1990 and 1991 were tested for mycoplasma-like organism (MLO) infection with a DNA probe that detected BBSD-associated MLO. The percentage of plants and the number of Scaphytopius spp. that were MLO-positive tended to follow the same bimodal distribution found in the population studies. BBSD transmission tests were performed with Scaphytopius spp. collected from the field. Stunt-related MLO transmission was achieved with S. magdalensis, S. acutus and 5. frontalis.     

Use of Polyclonal Antibodies to Identify Mycoplasma-like organisms (MLOs) From the Sudan and from Thailand

Rabbit polyclonal antibodies prepared against faba bean phyllody MLO from the Sudan reacted with its homologous antigen and with extracts of Catharanthus roseus experimentally infected with the same or a related MLO from Crotalaria saltiana showing symptoms of phyllody disease, as well as with extracts of naturally MLO-infected C. saltiana growing in the field in the Sudan. The antibodies also reacted positively with extracts of C. roseus experimentally infected with Crotalaria juncea phyllody MLO and soybean phyllody MLO from Thailand. Polyclonal antibodies prepared against an MLO associated with witches' broom disease in C. juncea reacted positively in ELISA tests with homologous antigen extracts from naturally infected C. juncea as well as with extracts from experimentally infected C. roseus and with extracts prepared from Sesamum indicum plants with phyllody symptoms growing in Thailand. There was no reaction between these antibodies and extracts from C. roseus plants infected with the MLOs associated with C. juncea phyllody or with soybean phyllody. No cross reactions were observed among the antigens and antibodies of the two MLO groups by immunoflorescence, ELISA or western blotting. However, the molecular weight of the principal protein antigen, determined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting was the same for both types of MLO. Serologically-similar MLOs thus occur in the Sudan and in Thailand, where they are associated with phyllody symptoms in C. saltiana and faba bean and with C. juncea and soybean, respectively. A second, serologically distinct MLO group was also found infecting C. juncea and S. indicum in Thailand but MLOs from this group have not yet been identified in crops from the Sudan.     

Amplification of plasmid DNA to detect plant pathogenic mycoplasmalike organisms

The polymerase chain reaction (PCR) was employed to develop a specific assay for plant pathogenic mycoplasmalike organisms (MLOs). A cloned fragment of a plasmid from a severe strain of western aster yellows (AY)-MLO was sequenced to identify oligonucleotide primers for PCR. Amplified DNA fragments of the predicted size were obtained from DNA extracted from plants and insects infected with pear decline MLO, beet leafhopper-transmitted virescence agent, elm yellows MLO and several AY-MLO strains. No amplification occurred from healthy leafhopper or plant DNA. The PCR-based assay was over 500 times more sensitive than a _tilized_tion-based assay which _tilized a cloned AY plasmid fragment as a probe. With the PCR-based assay, MLOs could be detected using DNA samples of leafhoppers that were only crushed and boiled in buffer. Amplification of the target DNA was confirmed by digestion of the PCR product with Mbo I which yielded predicted sized fragments for all MLO strains except Bradford AY and eastern AY. Sequencing the PCR product from elm yellows and eastern AY-MLOs revealed greater than 90% homology, and the failure to restrict the PCR product with Mbo I was due to a single base change in the restriction endonuclease site. The ability of the assay to detect a wide range of MLOs with minimal sample preparation and high sensitivity will be useful in epidemiological studies of MLO-caused diseases.     

Pathogenicity and effects on transmission of a mycoplasmalike organism of a transovarially infective bacterium on the leafhopper Euscelidius variegatus (Homoptera: Cicadellidae)

A transovarially transmitted, Gram-negative bacterium (BEV) reduced the median longevity of congenitally infected Euscelidius variegatus 54%, compared to uninfected controls of the same age and bred from the same parental stock. Mean fecundity was reduced over 80%, and nymphal development took almost 50% longer. Infected surviving adults of both sexes, however, weighed significantly more than uninfected controls. Infection of E. variegatus with BEV significantly reduced the transmission of the plant pathogenic mycoplasmalike organism causing X-disease in celery. The pathological effects of the BEV bacterium on its leafhopper host imply that infection by means other than transovarial transmission is necessary for the bacterium to persist in nature.     

Increased survival of Dalbulus maidis, a specialist on maize, on non-host plants infected with mollicute plant pathogens

The leafhopper Dalbulus maidis DeLong & Wolcott survived significantly longer on aster, Callistephus chinensis Nees, infected with any one of 3 strains of aster yellows (AY) mycoplasma-like organism (MLO) than on healthy asters. After 7 or more days on AY-diseased aster, females were conditioned to survive longer on healthy asters than were leafhoppers of the same age previously exposed only to maize. Females were also conditioned to survive longer on healthy aster by prior exposure to AY-MLO-infected celery (Apium graveolens L.). Males were not so conditioned. Leafhoppers injected with infectious extracts of AY-MLO dit not live longer on aster nor transmit the AY-MLO to aster. Conditioning on AY-diseased aster did not cause D. maidis to transmit AY-MLO and did not interfere with the transmission to maize of the mollicute (Spiroplasma kunkelii Whitcomb et al.) that causes corn stunt disease. Spiroplasma citri Saglio et al. infection of aster but not of turnip (Brassica rapa L.), Plantago major L. or periwinkle (Catharanthus roseus (L.)), improved the longevity of D. maidis on these plants and conditioned leafhoppers for enhanced subsequent survival on healthy asters.

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Résumé La cicadelle du maïs, D. maidis à survécu significativement plus longtemps sur aster (Callistephus chinensis Nees) contaminé par l'une des trois souches de l'agent mycoplasmique (MLO) de la jaunisse de l'aster (AY), que sur des asters sains. Après 7 jours ou plus sur des asters AY-MLO, les femelles (et non les mâles) étaient conditionnées de telle sorte qu'elles survivaient plus longtemps sur asters sains que des femelles du même âge précédemment sur maïs. Sur des céleris (Apium graveolens) atteints de AY, D. maidis a survécu plus longtemps que sur céleri sain, mais moins que sur aster AY. Des extraits, contaminés par AY injectés à D. maidis n'ont pas augmenté sa longévité sur aster, ni provoqué la transmission de AY-MLO. Le conditionnement sur aster AY n'a pas entrainé la transmission de AY-MLO par D. maidis, et n'a pas interferé avec son aptitude à transmettre l'agent du nanisme du maïs, Spiroplasma kunkelii. S. citri a aussi conditionné D. maidis sur aster, mais a été sans effet sur navet (Brassica rapa), sur plantain (Plantago major) et sur pervenche (Vinca).

     

Continuous rearing of the aster leafhopper, Macrosteles fascifrons, on a chemically defined diet

A holidic diet for feeding the aster leafhopper, Macrosteles fascifrons, was formulated. The amino acids, B-vitamins, and sucrose are less concentrated than in aphid diets. Cholesterol, at 5 mg/ml, is required for the last ecdysis. Although leafhoppers reared on this diet have poorer survival and shorter life span than those reared on plants, they produce more progeny. Leafhoppers reared on this diet have completed the ninth generation and the culture is still thriving.     

Etiology and Transmission of Sesame Phyllody in Iran

Phyllody is a destructive disease of sesame (Sesamum indicum L.) in Iran. The major symptoms of the disease are floral virescence, phyllody and proliferation. Other symptoms which sometimes accompany the disease are yellowing, cracking of seed capsules, germination of seeds in the capsules and formation of dark exudates on the foliage. Light microscopy of hand-cut sections of sesame and colza (Brassica napus L. cv. Oro) stems treated with Dienes' stain showed blue areas in the phloem region of phyllody infected plants. Mycoplasma-like bodies were found in the sieve cells of infected sesame stems when thin sections were examined m an electron microscope. Sesame phyllody was successfully transmitted from sesame to sesame by grafting. Among various leafhoppers collected in sesame fields only Neoaliturus haematoceps transmitted the disease. This is the first report on the identification of a Mycoplasma-like organism (MLO) as the cause of sesame phyllody and N. haematoceps as an MLO vector in Iran. In host range studies using the leafhopper vector, only B. napus cv. Oro, Lepidium sativum, Catharanthus roseus, Lactuca sp. and Portulaca oleracea, but not 17 other species, developed symptoms. The species of vector and host range of MLO indicate that sesame phyllody in Iran is different from that reported in India and Upper Volta.     

Comparison of CEL I gene expression and mismatch-cleavage activity in some Apiaceae plants

The most familiar enzyme in mutation screening through heteroduplex analysis, CEL I, has been isolated from celery of the Apiaceae family. In this study, in a search for new sources with the same or better enzymatic activity, we studied the mismatch-cleavage activity of plant juice extracts from several Apiaceae plants (celery, carrot, coriander, parsley, dill, and fennel). This study was then followed by investigation of the level of CEL I gene expression in these plants. Mismatch-cleavage activity of fennel and dill juice extracts was lower than that of celery juice extract, and levels of CEL I mRNA expression in these plants were substantially higher than in celery. In contrast, the ability of juice extract from a local cultivar of parsley to cleave heteroduplex DNA substrates was clearly more than that of celery juice extract, whereas the level of CEL I gene expression in parsley was obviously lower than in celery. We concluded that there are multiple mismatch-cleaving enzymes collaborating in digestion of heteroduplex DNA substrates by plant juice extracts.     

Aphid-injection experiments with carrot mottle virus and its helper virus, carrot red leaf

Carrot mottle virus (CMotV) and its helper virus, carrot red leaf (CRLV), were not transmitted by aphids (Cavariella aegopodii) that had fed through membranes on, or had been injected with, sap from mixedly infected chervil plants or partially purified preparations of CMotV. However, the viruses were transmitted by recipient aphids injected with haemolymph from donor aphids that had fed on mixedly infected plants but not by a second series of recipients injected with haemolymph from the first series. Some of the first series of recipients transmitted both viruses for up to 11 days but others transmitted erratically and many lost ability to transmit after a few days. The results confirm that both viruses are circulative but provide no evidence for multiplication in the vector. Non-viruliferous aphids, or aphids that had acquired CRLV by feeding, did not transmit CMotV when they were injected with haemolymph from aphids that had fed on a source of CMotV alone, confirming that they can only transmit CMotV when they acquire it from a mixedly infected plant. When extracts from donor aphids were treated with ether before injection, recipient aphids transmitted both CRLV and CMotV, although the infectivity of CMotV grown in Nicotiana clevelandii in the absence of CRLV is destroyed by ether treatment. CMotV particles acquired by aphids from mixedly infected plants therefore differed in some way from those in singly infected plants. A plausible explanation of these results, and of the dependence of CMotV on CRLV for aphid transmission, is that doubly infected plants contain some particles that consist of CMotV nucleic acid coated with CRLV protein.     

The sensory systems and feeding behavior of leafhoppers. II. A comparison of the sensillar morphologies of several species (Homoptera: Cicadellidae)

The ultrastructure of the sensilla, and other structures, within the precibaria of eight species from three subfamilies of leafhoppers (Homoptera: Cicadellidae) were examined with scanning electron microscopy. The types and grouping of the 20 precibarial sensilla in seven of these species were similar to those observed previously in Macrosteles fascifrons Stål. Oncometopia nigricans (Walker) also displayed similar sensilla groups; however, it had 30 sensilla. The species examined differed chiefly in the exact location and arrangement of the sensilla. The possible significance of the differences relative to leafhopper feeding is discussed. The precibarial chemosensilla may provide chemosensory evaluation of fluid in the food canal and precibarium prior to ingestion or egestion.     

Mycoplasma-like bodies inSolanum laciniatum plants infected with potato witches’ broom disease

Mycoplasma-like organisms (MLO) spread from the infectious grafts intoSolanum laciniatum Ait. stock plants relatively slowly. MLO were present in all sprouts ofS. laciniatum four weeks after grafting, but the infected plants remained under glasshouse conditions mostly symptomless and flowered normally and formed fruits like healthy plants. The growth of plants with infectious tomato grafts was identical with the controls but that of plants with infectious tobacco (Nicotiana glauca Grah.) grafts was expressively stimulated. The first flower symptoms appeared onS. laciniatum plants with tomato grafts after five and half months and on.S. laciniatum plants with tobacco grafts after seven months of graft symbiosis. Electron micrographs of ultrathin sections showed the presence of MLO in sieve tubes of potiols and midribs of the infected but symptomless plants. In the phloem parenchyma cells of the witches’ broom diseased plants, highly ordered crystals were occasionally found lying in a microbody surrounded by a membrane. The possible reasons of the disease latency are discussed.     

The sensory systems and feeding behavior of leafhoppers. I. The aster leafhopper,Macrosteles fascifrons stål (homoptera,cicadellidae)

The ultrastructure of the sensilla, and other structures, within the stylets and precibarium of Macrosteles fascifrons were examined by transmission and scanning electron microscopy. Precibarium is a new term, defined here, for the canal that precedes the cibarium inside the leafhopper head. Within the precibarium are found 20 chemosensilla and a previously undescribed structure, the precibarial valve. Twelve mechanosensilla, three in each stylet, are found within the maxillary and mandibular stylets. The relationship between all of these structures and feeding by the insect is detailed in a feeding mechanism hypothesis. It is concluded that leafhoppers (and probably all homopterans) utilize the precibarial chemosensilla alone for gustatory discrimination, the stylet sensilla for proprioception, and the precibarial valve for regulation of fluid uptake and compartmentalization of the sensilla.     

The presence of <Emphasis Type="Italic">Zygina</Emphasis> sp. and <Emphasis Type="Italic">Puccinia myrsiphylli</Emphasis> reduces survival and influences oviposition of <Emphasis Type="Italic">Crioceris</Emphasis> sp.

A leaf beetle, Crioceris sp. (Coleoptera: Chrysomelidae), was introduced into Australia as a biological control agent of bridal creeper (Asparagus asparagoides L. Druce) during October 2002. Rearing of Crioceris sp. is labour intensive therefore all releases of Crioceris sp. have been under 1000 individuals, which may be too low to ensure establishment if high mortality and high competition with other agents occurs. The aim of this study is to understand how the presence of two well-established biocontrol agents, a rust fungus (Puccinia myrsiphylli (Thuem) Wint [Basidiomycota: Uredinales]) and a leafhopper (Zygina sp. [Hemiptera: Cicadellidae]), might influence Crioceris sp. establishment. Crioceris sp. neonate larvae were placed on bridal creeper plants with or without the leafhopper and/or rust. The number of larvae that pupated was reduced by 38 and 65% in the presence of the rust fungus and leafhopper, respectively and by 45% in the presence of both agents. As the area infected by the rust increased the area damaged by the leafhopper decreased. The rust appeared to be negatively impacted by the presence of the leafhopper. In a second experiment, female Crioceris sp. adults were given a choice between uninfested bridal creeper plants and those infested with the rust or the leafhopper. The females preferred to lay their eggs on plants without leafhoppers but did not seem to be deterred by the presence of the rust. Consequently, the performance and impact of Crioceris sp. on bridal creeper may be reduced if populations overlap with the other biocontrol agents in the field.     

Detection of anti-tuberculosis activity in some folklore plants by radiometric BACTEC assay

Aims: The anti‐tubercular drugs are less effective because of the emergence of multi‐drug resistant (MDR) and extensively drug resistant (XDR) strains of M. tuberculosis, so plants being an alternative source of anti‐microbial compounds. The aim of this study was to investigate anti‐tuberculosis potential of the plants using Mycobacterium smegmatis as a rapid screening model for detection of anti‐mycobacterial activity and further to evaluate the active plants for anti‐tuberculosis activity against M. tuberculosis using radiometric BACTEC assay. Methods and Results: The 15 plants were screened for anti‐mycobacterial activity against M. smegmatis by the disk diffusion assay. The ethanolic extracts of Mallotus philippensis, Vitex negundo, Colebrookea oppositifolia, Rumex hastatus, Mimosa pudica, Kalanchoe integra and Flacourtia ramontchii were active against M. smegmatis in primary screening. The anti‐tuberculosis potential was identified in the leaves extracts of Mallotus philippensis by radiometric BACTEC assay. The ethanolic extract of M. philippensis showed anti‐tuberculosis activity against virulent and avirulent strains of M. tuberculosis H₃₇Rv and M. tuberculosis H₃₇Ra with minimum inhibitory concentration 0·25 and 0·125 mg ml^?1, respectively. The inhibition in growth index values of M. tuberculosis was observed in the presence of ethyl acetate fraction at a minimum concentration of 0·05 mg ml^?1. Conclusion: We found that BACTEC radiometric assay is a valuable method for detection of anti‐tuberculosis activity of the plant extracts. The results indicate that ethanolic extract and ethyl acetate fraction of M. philippensis exhibited significant anti‐mycobacterial activity against M. tuberculosis. Significance and Impact of the Study: These findings provide scientific evidence to support the traditional medicinal uses of M. philippensis and indicate a promising potential of this plant for the development of anti‐tuberculosis agent.