Cassava whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in East African farming landscapes: a review of the factors determining abundance.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a pest species complex that causes widespread damage to cassava, a staple food crop for millions of households in East Africa. Species in the complex cause direct feeding damage to cassava and are the vectors of multiple plant viruses. Whilst significant work has gone into developing virus-resistant cassava cultivars, there has been little research effort aimed at understanding the ecology of these insect vectors. Here we assess critically the knowledge base relating to factors that may lead to high population densities of sub-Saharan African (SSA) B. tabaci species in cassava production landscapes of East Africa. We focus first on empirical studies that have examined biotic or abiotic factors that may lead to high populations. We then identify knowledge gaps that need to be filled to deliver sustainable management solutions. We found that whilst many hypotheses have been put forward to explain the increases in abundance witnessed since the early 1990s, there are little published data and these tend to have been collected in a piecemeal manner. The most critical knowledge gaps identified were: (i) understanding how cassava cultivars and alternative host plants impact population dynamics and natural enemies; (ii) the impact of natural enemies in terms of reducing the frequency of outbreaks and (iii) the use and management of insecticides to delay the development of resistance. In addition, there are several fundamental methodologies that need to be developed and deployed in East Africa to address some of the more challenging knowledge gaps.

Skeletal overgrowth and deafness in mice lacking fibroblast growth factor receptor 3.

Fibroblast growth factor receptor 3 (Fgfr3) is a tyrosine kinase receptor expressed in developing bone, cochlea, brain and spinal cord. Achondroplasia, the most common genetic form of dwarfism, is caused by mutations in FGFR3. Here we show that mice homozygous for a targeted disruption of Fgfr3 exhibit skeletal and inner ear defects. Skeletal defects include kyphosis, scoliosis, crooked tails and curvature and overgrowth of long bones and vertebrae. Contrasts between the skeletal phenotype and achondroplasia suggest that activation of FGFR3 causes achondroplasia. Inner ear defects include failure of pillar cell differentiation and tunnel of Corti formation and result in profound deafness. Our results demonstrate that Fgfr3 is essential for normal endochondral ossification and inner ear development.

On species delimitation, hybridization and population structure of cassava whitefly in Africa.

The Bemisia cassava whitefly complex includes species that cause severe crop damage through vectoring cassava viruses in eastern Africa. Currently, this whitefly complex is divided into species and subgroups (SG) based on very limited molecular markers that do not allow clear definition of species and population structure. Based on 14,358 genome-wide SNPs from 62 Bemisia cassava whitefly individuals belonging to sub-Saharan African species (SSA1, SSA2 and SSA4), and using a well-curated mtCOI gene database, we show clear incongruities in previous taxonomic approaches underpinned by effects from pseudogenes. We show that the SSA4 species is nested within SSA2, and that populations of the SSA1 species comprise well-defined south-eastern (Madagascar, Tanzania) and north-western (Nigeria, Democratic Republic of Congo, Burundi) putative sub-species. Signatures of allopatric incipient speciation, and the presence of a 'hybrid zone' separating the two putative sub-species were also detected. These findings provide insights into the evolution and molecular ecology of a highly cryptic hemipteran insect complex in African, and allow the systematic use of genomic data to be incorporated in the development of management strategies for this cassava pest.

Electron-opaque fibrils and granules in and between the cell walls of higher plants.

The components of higher-plant cell walls which become electron-opaque after staining with ruthenium-osmium were studied by electron microscopy. A fibrillar material which absorbs this stain is a major wall constituent in the root epidermal cells of carrot and morning glory. In both form and size, these fibrils resemble those found on the surface of suspension-cultured cells of the same species Some cells of woody species show an irregular distribution of electron-opaque material in the cell wall matrix and middle lamella. This material, which has an amorphous appearance with many electron stains, is shown by ruthenium-osmium staining to be an aggregate of discrete granules, 150-220 A in diameter. These observations are not consistent with the concept of the cell wall matrix and middle lamella as an amorphous, uniform gel

Lignofibrils on the external cell wall surface of cultured plant cells.

Small strands and bundles of strands extend from the outside surface of suspension-cultured cells of Daucus, Ipomoea, and Phaseolus into the medium. This fibrous cell coat is present in all samples from various growth stages but appears to increase in quantity in the order Ipomoea < Phaseolus < Daucus. The bundles are often many microns in length and display great variation in frequency, size, and form. Identification of the composition of the strands and bundles as lignin is consistent with the following observations: alkaline nitrobenzene oxidation of the strands to compounds which resemble monomers of wood lignin; resistance of the strands to pronase, trypsin, pectinase, and lipase; strong irreversible adsorption of heavy metals; deposition of silver granules by treatment with silver nitrate-hexamine reagent; extraction of the bundles with aqueous dioxane (Björkman procedure); presence in quantity of a structured form of Klason lignin; and existence of material giving a positive test with the Wiesner reagent. Large individual strands (lignofibrils) from Phaseolus show the form of a flat ribbon with very thin branches at irregular intervals. This form does not vary with preparatory techniques, although its electron opacity does. Intercellular spaces display considerable structure and sometimes contain sheets of fibrillar material merging with both the middle lamella between the cells and the surface bundles facing the medium. These sheets are probably another form of association of the lignofibrils. It is suggested that natural fibrous lignin may be a much commoner component of plant tissue than suspected hitherto.

Re-emergence of Cassava Brown Streak Disease in Uganda.

During November 2004, veinal chlorosis on mature cassava leaves, typical of cassava brown streak disease (CBSD), was observed at Mukono in central Uganda. Five out of 11 cultivars at the site showed CBSD symptoms (incidence range 4 to 64%). In a survey of farmers' fields, CBSD was observed in Wakiso and Mukono districts. Incidence of cassava mosaic disease was also recorded and averaged 60% for landraces (range 16.7 to 100%) and 20% for resistant varieties (range 0 to 65%). Leaf samples of plants with CBSD symptoms produced an amplicon of 222 bp using reverse transcription-polymerase chain reaction with primers that amplify a fragment of the coat protein (CP) gene of Cassava brown streak virus. Sequence comparisons based on the amplified CP gene fragment indicated that the isolates have 77 to 82.9% nucleotide and 43.9 to 56.8% amino acid identity with those from Mozambique and Tanzania. There was 95.9 to 99.5% nucleotide and 85.1 to 90.5% amino acid identity among the Ugandan isolates. These results confirm the re-emergence of CBSD in Uganda after it was first observed in the 1930s in cassava introduced from Tanzania and controlled by eradication. Prior to this report, CBSD was known to be restricted to the coastal lowlands of East Africa.

Predicting the thermal inactivation of bacteria in a solid matrix: simulation studies on the relative effects of microbial thermal resistance parameters and process conditions.

A combined mathematical model for predicting heat penetration and microbial inactivation in a solid body heated by conduction was tested experimentally by inoculating agar cylinders with Salmonella typhimurium or Enterococcus faecium and heating in a water bath. Regions of growth where bacteria had survived after heating were measured by image analysis and compared with model predictions. Visualisation of the regions of growth was improved by incorporating chromogenic metabolic indicators into the agar. Preliminary tests established that the model performed satisfactorily with both test organisms and with cylinders of different diameter. The model was then used in simulation studies in which the parameters D, z, inoculum size, cylinder diameter and heating temperature were systematically varied. These simulations showed that the biological variables D, z and inoculum size had a relatively small effect on the time needed to eliminate bacteria at the cylinder axis in comparison with the physical variables heating temperature and cylinder diameter, which had a much greater relative effect.

Understanding reconstructed Dante spectra using high resolution spectroscopy.

The Dante is an 18 channel filtered diode array used at the National Ignition Facility (NIF) to measure the spectrally and temporally resolved radiation flux between 50 eV and 20 keV from various targets. The absolute flux is determined from the radiometric calibration of the x-ray diodes, filters, and mirrors and a reconstruction algorithm applied to the recorded voltages from each channel. The reconstructed spectra are very low resolution with features consistent with the instrument response and are not necessarily consistent with the spectral emission features from the plasma. Errors may exist between the reconstructed spectra and the actual emission features due to assumptions in the algorithm. Recently, a high resolution convex crystal spectrometer, VIRGIL, has been installed at NIF with the same line of sight as the Dante. Spectra from L-shell Ag and Xe have been recorded by both VIRGIL and Dante. Comparisons of these two spectroscopic measurements yield insights into the accuracy of the Dante reconstructions.

Nucleotide sequencing, whitefly transmission, and screening tomato for resistance against two newly described begomoviruses in bangladesh.

ABSTRACT The molecular diversity of Tomato leaf curl viruses (ToLCVs), from the two main tomato growing areas of Jessore and Joydebpur, Bangladesh, was investigated. The viral DNA was amplified from tomato plants exhibiting mild and severe symptoms by polymerase chain reaction, and the complete genomes of the ToLCVs were sequenced. An isolate of the bipartite Tomato leaf curl New Delhi virus-Severe (ToLCNDV-Svr) was associated with the severe symptom phenotype from Jessore (ToLCNDV-Svr[Jes]). A previously undescribed monopartite virus, designated Tomato leaf curl Joydebpur virus-Mild (ToLCJV-Mld), was sequenced from plants showing mild symptoms. ToLCNDV-Svr[Jes] was most closely related to ToLCNDV-[Lucknow] at 95.7% nucleotide (nt) identity and Tomato leaf curl Gujarat virus-[Varanasi] at 90.6% nt identity, based on DNA-A and -B component sequences. ToLCJV-Mld was similar to Pepper leaf curl Bangladesh virus at 87.1% DNA-A nt identity. Identification of ToLCNDV-Svr[Jes] and ToLCJV-Mld was in addition to the previously described Tomato leaf curl Bangladesh virus, with which they shared 73.2 and 86.0% DNA-A nt identities, thus demonstrating the existence of at least three distinct viruses infecting tomato in Bangladesh. Nucleotide identities and placement in phylogenetic trees suggested that the three ToLCVs may have had different evolutionary pathways. The whitefly, Bemisia tabaci, transmitted the viruses of this study equally efficiently. Four tomato cultivars (TLB111, TLB130, TLB133, and TLB182) resistant/ tolerant to South Indian ToLCV were screened against the Bangladesh ToLCVs in 2003-04. Although challenged by diverse viruses and potentially mixed infections, disease incidence remained low (6 to 45%) in the resistant cultivars compared with local cultivars (68 to 100%).

First Report of Tomato leaf curl New Delhi virus Infecting Tomato in Bangladesh.

Tomato is an important cash crop for resource-poor farmers and accounts for 20% of the 2 million t of vegetables grown annually in Bangladesh. Tomato cultivation is affected by Tomato leaf curl virus (ToLCV), which can cause as much as 100% yield loss. Plants exhibiting typical ToLCV disease symptoms of yellowing, severe leaf curling, and stunting were collected at Jessore, Bangladesh during September 2003. The putative virus was transmitted from tomato to tomato by the whitefly Bemisia tabaci. In two separate experiments, 100% transmission was achieved by using 10 viruliferous B. tabaci adults for each of the 20 test plants that was confirmed by comparing the symptoms on test and virus source plants. Total DNAs were extracted from the symptomatic leaves, and the putative viral genomes were amplified by polymerase chain reaction by using the Deng A and B primers (1). Sequences generated from these primers were used to design virus-specific primers that were used to obtain complete viral sequences. Full-length DNA-A (2,740 nt; GenBank Accession No. AJ875157) and DNA-B (2,688 nt; GenBank Accession No. AJ875158) sequences of a bipartite Tomato leaf curl New Delhi virus from Jessore (ToLCNDV-[Jes]) were obtained, which were most similar to the corresponding sequences of ToLCNDV-(Lucknow) (GenBank Accession No. Y16421) at 95.7% and Tomato leaf curl Gujarat virus-(Varanasi) (Gen-Bank Accession No. AY190291) at 90.6% nt identities, respectively. DNA-A sequences had only 73.2% nt identity with the previously reported monopartite Tomato leaf curl Bangladesh virus (GenBank Accession No. AF188481) (2), confirming the occurrence of mono- and bipartite bego-moviruses in Bangladesh. The virus diversity poses a challenge for ToLCVD management in Bangladesh. References: (1) D. Deng et al. Ann. Appl. Biol. 125:327, 1994. (2) S. K. Green et al. Plant Dis. 85:1286, 2001.

Variability of the iron, copper and mercury contents of individual red blood cells.

The relative iron, copper and mercury contents of individual, isolated erythrocytes from eight people were determined by analytical electron microscopy. The variation in iron content between erythrocytes of the same sample is more than six times, for copper content more than tem times and for mercury more than five times. Similar variations were observed for 1-day-old chick defintive erythrocytes and for 4-day-old chick embryo primitive erythrocytes. The range of variation does not depend greatly, it at all, on the age of the erythroyctes or the tissue of origin. There is little or no correlation between the variation of iron contnet and that of copper. The cause of the wide variation of metallic ion contnt among erythroyctes is not yet known.

Radionuclide cerebral angiographic evaluation of a diploic extracranial meningioma: case report.

A case of a diploic extracranial meningioma whose true vascularity was more significantly defined by radionuclide angiography than by selective contrast angiography is presented. The radionuclide angiogram clearly demonstrated large venous channels draining the tumor.

Co-adaptation between cassava mosaic geminiviruses and their local vector populations.

Four cassava mosaic geminivirus (CMG) isolates; African cassava mosaic virus from Namulonge, Uganda (ACMV-[Nam]), East African cassava mosaic virus from Mtwara, Tanzania (EACMV-[Mtw]), EACMV-Uganda from Namulonge (EACMV-UG[Nam]) and Indian cassava mosaic virus from Trivandrum, India (ICMV-[Tri]) were compared for their ability to be transmitted by four colonies of cassava whitefly, Bemisia tabaci (Gennadius), collected from Namulonge (NAM), Mtwara (MTW), Kumasi (KUM) and Trivandrum (TRI). With 20 adult whiteflies per test plant, the CMGs from Africa were transmitted by all three of the African B. tabaci populations to 60-79% of the target plants. Indian cassava B. tabaci transmitted ICMV-[Tri] to 89% of the target plants. In contrast, Indian cassava B. tabaci transmitted EACMV-[Mtw] and Tanzanian cassava B. tabaci transmitted ICMV-[Tri] less than a tenth as often, even when using 50 adults per plant and with increased acquisition and inoculation access periods. The complete coat protein genes of the CMGs had sequences typical of their source viruses, the major differences occurring between those originating from India and Africa. Symptom severity of the CMGs was quantified precisely by both visual assessment and image analysis techniques. EACMV-[Mtw] and ACMV-[Nam] were the most and least damaging isolates with 15.4 and 10.0% of the leaf area diseased, respectively. In these tests the transmission frequency was not linked to symptom severity in the source plants. These data support the hypothesis that virus-vector co-adaptation exists in the cassava mosaic disease (CMD) pathosystem and the results are discussed in relation to CMD epidemiology.

Use of 2% propofol to produce diurnal sedation in critically ill patients.

OBJECTIVE: The assessment of propofol to produce diurnal sedation in critically ill patients. DESIGN: Prospective clinical study. SETTING: Intensive Care Unit, University Hospital. PATIENTS AND PARTICIPANTS: Thirty consecutive patients admitted to the Intensive Care Unit older than 18 years who were expected to be sedated for more than 50 h. INTERVENTIONS: The patients were randomised into two groups. All received sedation with a constant background infusion of morphine and a variable infusion rate of propofol, which was altered hourly to maintain the intended sedation score. The first group received constant light sedation (CLS) over 50 h aiming for a Ramsay score of 2-3. The second group received CLS between 0600 h and 2200 h and additional night sedation (ANS) with propofol between 2200 h and 0600 h, aiming for a sedation score of 4-5. MEASUREMENTS AND RESULTS: Patients were studied for 50 h from 1800 h on the first day of admission. Recordings of heart rate, blood pressure, sedation scores and propofol and morphine infusion rates were made hourly. An APACHE II score was recorded for each patient. Sedation scores were analysed by blind visual assessment and cosinor analysis, which is used in chronobiology to examine the correlation of data with a cosine curve. Patients in the ANS group had significantly better rhythmicity of sedation levels using cosinor analysis (r = 26% v 8%) p < 0.01. There was no difference between the CLS and ANS groups with respect to age, sex or APACHE II scores. Nine out of 15 patients in the ANS group achieved diurnal sedation. Three patients in the CLS group showed diurnal rhythmicity of sedation, which can be attributed to natural sleep, and had a median APACHE II score of 12. Five patients in the CLS group and three in the ANS group showed a deep constant sedation pattern. They had high APACHE II scores (median 21.5) and an obtunded conscious level on admission due to severe sepsis. CONCLUSION: Propofol can safely provide diurnal sedation in the critically ill when titrated against the Ramsay score. Sedation levels cannot be manipulated in some severely ill patients.

Shaping attitudes to postoperative pain relief: the role of the acute pain team.

Postoperative pain relief is often inadequate. Ignorance and misconceptions about opioids by ward staff contribute to this poor management. The introduction of acute pain teams has done much to improve pain relief for patients. It may also have contributed to changes in attitudes and knowledge of medical and nursing staff. We questioned 48 doctors and nurses on their knowledge and beliefs about postoperative pain relief. Staff members were questioned on two units, one with access to an acute pain team and one without. Over half those on the unit using traditional postoperative care thought patients did not receive adequate pain relief (58%). In comparison, only one respondent from the unit with the pain team thought this was the case (P < 0.001). More staff members that had experience of patient-controlled analgesia (PCA) were optimistic about its benefits than those in the unit with no experience; they were also less concerned about possible side effects. Only one respondent on the unit using PCA thought it carried a risk of drug dependence, compared to over half (55%) of those on the unit with no experience in this technique (P < 0.001). Over two-thirds of staff familiar with PCA thought nursing workload had decreased. Acute pain teams have an important role in educating ward staff. The impact of establishing such teams on staff knowledge and attitudes needs further study to ensure that they can carry out this role most effectively.

Valproate-induced limb malformations in mice associated with reduction of intracellular pH.

Valproic acid (VPA) is a commonly used antiepileptic agent that recently has been found useful in the treatment of affective disorders and prophylaxis of migraine. VPA induces congenital malformations, especially spina bifida, in the offspring of women treated with this agent during early pregnancy. The mechanism by which VPA induces abnormal development remains unknown despite many studies in experimental animals in which VPA causes malformations similar to those seen in human infants. Because of its chemical structure as a weak organic acid and its capability to induce postaxial forelimb ectrodactyly in C57BL/6 mice, we postulated that VPA acts to perturb limb morphogenesis by reducing embryonic intracellular pH (pHi). We administered VPA, 200 to 400 mg/kg, to C57BL/6 mice on day 9 of gestation. A dose-dependent incidence of postaxial forelimb ectrodactyly was observed. Forelimb bud pHi was estimated by computer-assisted image analysis from the transplacental distribution of 14C-DMO. At the highest doses, 300 and 400 mg/kg, a decrease of pHi of 0.2 to 0.3 pH units was observed uniformly throughout the limb bud 1 h after VPA treatment. None of these changes were seen after treatment with 2-en VPA, a nonteratogenic analog of VPA. Furthermore, the capability of VPA to induce postaxial forelimb ectrodactyly was greatly enhanced by coadministration of agents that inhibit pHi regulatory processes. These data support the hypothesis that VPA-induced postaxial ectrodactyly in murine fetuses can be attributed to reduction in limb bud pHi.

Mathematical models of host plant infection by helper-dependent virus complexes: why are helper viruses always avirulent?

ABSTRACT Interactions between viruses in plants are common, and some viruses depend on such interactions for their survival. Frequently, a virus lacks some essential molecular function that another provides. In "helper-dependent" virus complexes, the helper virus is transmitted independently by a vector, whereas the dependent virus depends on molecular agents associated with the helper virus for transmission by a vector. A general mathematical model was developed of the dynamics of host plant infection by a helper-dependent virus complex. Four categories of host plants were considered: healthy, infected with helper virus alone, infected with dependent virus alone, and infected with both viruses. New planting of the host crop was constrained by a maximum abundance due to limitation of the cropping area. The ratio of infection rate to host loss rate due to infection is proposed as an important epidemiological quantity, A, that can be used as a measure of the mutual adaptation of the virus and host. A number of alternative equilibria of host infection could occur and were determined exclusively by parameter values; it was informative to display their distribution in the parameter plane: (1/A)(helper) versus (1/A)(dependent). A simple analysis of the distribution of the final equilibria illustrated that the dependent virus could affect the survival of the helper virus, so facilitation between the two can be reciprocal. The distribution of the final equilibria also indicated that a well-adapted helper virus increases the opportunity for a dependent virus to evolve and survive, and the model, therefore, explains why infection with a helper virus usually causes no or little damage to plants, whereas infection with a dependent virus or mixed infection with both often causes very severe damage.

First Report of a Distinct Begomovirus Infecting Cassava from Zanzibar.

In 1998, cassava plants exhibiting extremely mild mosaic disease symptoms were collected from Uguja Island, Zanzibar. Total DNAs extracted from symptomatic leaves did not produce diagnostic PCR bands using primers specific to known Cassava mosaic viruses (genus Begomovirus, family Geminiviridae) (2). Degenerate primer pair A/B (1), however, produced a 564-nucleotide (nt) band in the common region of DNA-A to the conserved amino acid sequence CEGPCKYG within the coat protein gene for begomoviruses. Virus-specific primers were designed and the begomoviral genome was amplified and cloned to obtain the full-length DNA-A (2,785 nt) (AF422174) and the partial DNA-B (2,403 nt) (AF422175) component comprising the 5' end of the BV1 open reading frame (ORF) to eight (A) of the conserved nanonucleotide TAATATTAC in the common region. Phylogenetic comparisons of DNA-A and -B sequences of the cassava begomivirus showed they were most closely related to East African cassava mosaic virus (EACMV) from Tanzania (Z83256, 83% identity) and Uganda (AF126805, 87% identity). Based on DNA-A and -B components, its closest relatives among African cassava mosaic virus strains were the Nigerian (X17095, 67% identity) and Ugandan (AF126800, 38% identity) isolates respectively. The number and arrangement of the viral ORF was identical to other bipartite begomoviruses from the Old World. Work is in progress to determine whether this begomovirus is a new strain of an existing EACMV or a new species. References: (1) D. Deng et al. Ann. App. Biol. 125:327, 1994. (2) X. Zhou et al. J. Gen. Virol. 78:2101, 1997.

First Report of the Bemisia tabaci B Biotype in India and an Associated Tomato leaf curl virus Disease Epidemic.

In May 1999, in the Kolar district of Karnataka State, Bemisia tabaci numbers on tomato increased by approximately 1,000-fold that observed previously (3). This was associated with an epidemic of severe tomato leaf curl disease that caused complete crop failure. DNAs extracted from 35 symptomatic tomato leaf samples collected within the epidemic region all gave the expected 500 to 600 bp amplicon with begomovirus-specific primers A/B (1). These primers amplify from the conserved nonanucleotide TAATATTAC in the common region of DNA-A to the conserved amino acid sequence CEGPCKYG within the coat protein gene. AluI and TaqI restriction patterns of all 35 polymerase chain reaction (PCR) products were identical. One PCR product from an epidemic (GenBank no. AF321929) and a non-epidemic (AF321930) site (Bangalore) were cloned and sequenced. The two 531-bp inserts showed 96% nucleotide identity to each other and 94% nucleotide identity to the equivalent region of Tomato leaf curl Bangalore virus (ToLCBV-Ban-4) (AF165098), suggesting that the epidemic was caused by an indigenous ToLCBV strain. Adult B. tabaci were collected from tomato plants at nine sites within the epidemic. DNA was extracted from 9 to 13 individuals per site and analyzed by RAPD-PCR using primers OpB20 and OpB11. Eighty to 100% of individuals per site had identical patterns to those of B biotype individuals from Israel and Florida, which were different to the patterns produced by the indigenous Indian B. tabaci. Adult B. tabaci from the epidemic and nonepidemic (Bangalore) regions were cultured separately on zucchini plants (n = 20) vars. Fordhook and Ambassador. Distinct silverleaf symptoms appeared in all plants fed on by the epidemic B. tabaci, but not on those fed on by the nonepidemic whiteflies. Irregular ripening of tomatoes was also a widespread problem in the epidemic area. Cytochrome oxidase I (COI) (720 bp) gene sequences were obtained for epidemic (AF321927) and nonepidemic (AF321928) B. tabaci, which had only 80% nucleotide identity to each other. A GenBank BLAST search showed that the former were most similar to B biotype whitefly from Israel (AF164667; 97%) and Texas (AF164675; 99%). The B biotype transmits Indian ToLCBV (2) and its introduction into India is of great concern as it is already associated with a devastating plant-disease epidemic. References: (1) D. Deng et al. Ann. App. Biol. 125:327, 1994. (2) P. F. McGrath and B. D. Harrison. Ann. App. Biol. 126:307, 1995. (3) H. K. Ramappa et al. Ann. App. Biol. 133:187, 1998.