Classical biological control of Aphis gossypii (Homoptera: Aphididae) with Neozygites fresenii (Entomophthorales: Neozygitaceae) in California cotton

In August 1994 and 1995 classical biological control releases were made in cotton in the San Joaquin Valley, California, with an Arkansas strain of the entomopathogenic fungus, Neozygites fresenii, a pathogen of the cotton aphid, Aphis gossypii. Pre-release samples in both years indicated that N. fresenii was not naturally present in A. gossypii populations in the San Joaquin Valley. Two release methods were compared: dried N. fresenii-infected cotton aphid “cadavers” and chamber inoculation of A. gossypii. Both methods were successful in introducing N. fresenii to cotton aphids in California; however, higher prevalence of fungal infection resulted with the cadaver treatments. N. fresenii persisted and spread in the aphid population until early October 1994 and late September 1995. The highest mean percentage infection in the cadaver treatment in 1994 reached a level (14%) considered imminent for epizootics (12–15%). The use of predator exclusion cages resulted in higher N. fresenii prevalences.     

Breeding Perennial Grain Crops

Referee: Ms. Peggy Wagoner, Rodale Institute, 611 Siegfriedale Road, Kutztown, PA 19530-9749 One-third of the planet's arable land has been lost to soil erosion in recent decades, and the pace of this degradation will increase as the limits of our food production capacity are stretched. The persistent problem of worldwide soil erosion has rekindled interest in perennial grain crops. All of our current grain crops are annuals; therefore, developing an array of new perennial grains - grasses, legumes, and others – will require a long-term commitment. Fortunately, many perennial species can be hybridized with related annual crops, allowing us to incorporate genes of domestication much more quickly than did our ancestors who first selected the genes. Some grain crops — including rye, rice, and sorghum — can be hybridized with close perennial relatives to establish new gene pools. Others, such as wheat, oat, maize, soybean, and sunflower, must be hybridized with more distantly related perennial species and genera. Finally, some perennial species with relatively high grain yields — intermediate wheatgrass, wildrye, lymegrass, eastern gamagrass, Indian ricegrass, Illinois bundleflower, Maximilian sunflower, and probably others — are candidates for direct domestication without interspecific hybridization. To ensure diversity in the field and foster further genetic improvement, breeders will need to develop deep gene pools for each crop. Discussions of breeding strategies for perennial grains have concentrated on allocation of photosyn-thetic resources between seeds and vegetative structures. However, perennials will likely be grown in more diverse agro-ecosystems and require arrays of traits very different from those usually addressed by breeders of annuals. The only way to address concerns about the feasibility of perennial grains is to carry out breeding programs with adequate resources on a sufficient time scale. A massive program for breeding perennial grains could be funded by diversion of a relatively small fraction of the world's agricultural research budget.     

Routine utilization of green fluorescent protein as a visual selectable marker for cereal transformation

Summary Development of new selectable markers is needed to increase the efficiency and flexibility of plant transformation, and to overcome drawbacks sometimes associated with use of existing markers. A useful alternative to chemical-based selection systems would be a system using visual screening to obtain transgenic lines. Investigations were carried out to determine if the green fluorescent protein (gfp) gene could be utilized alone as a visual screenable marker to produce stably transformed, fertile oat plants. Twelve experiments were conducted in which gfp-based selection was utilized to obtain routinely stable transgenic lines in oat. A synthetic gfp gene under the control of the maize ubiquitin promoter was delivered into embryogenic oat callus via microprojectile bombardment. Cell clusters (1–3 mm) expressing gfp were visually identified using epifluorescence microscopy and physically isolated approximately 3 wk post-bombardment. Fertile, gfp-expressing T0 plants were regenerated from 78% of the glowing cell sectors placed on regeneration medium. T0 plants from 55% of the events produced gfp-expressing progeny in a 3∶1 Mendelian ratio. Southern blot and PCR analysis confirmed transgene integration and transmission to progeny. Expression of gfp did not reduce plant growth or fertility. Transgene copy number and integration patterns were similar to those in transgenic plants derived from chemical-based selection systems. The mean transformation frequency, based on fertile events obtained per bombarded plate, was 1.8%. Over 180 independent transgenic oat lines were produced, to date, using only visual screening for expression of gfp, demonstrating efficiency and repeatability of the selection system. Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the University of Wisconsin or the US Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable.     

杀虫剂及不同地区寄主植物对棉蚜酯酶的影响

通过滤纸酯酶反应法比较 4种有机磷杀虫剂对北京和山东高密地区取食不同寄主植物的棉蚜β-乙酸萘酯 (β -NA)酯酶的抑制作用。结果表明高密地区不同寄主植物上的棉蚜 β -NA酯酶活力高的个体多于北京地区 ,同时辛硫磷对高密地区不同寄主植物上的棉蚜 β -NA酯酶的抑制作用亦高于北京地区 ,而久效磷和DDVP对两地区不同寄主植物上的棉蚜种群中 β-NA酯酶的抑制作用相似。     

Morphological and morphometrical analysis of Heterodera spp. populations in Jordan

Phenotypic diversity of five Jordanian populations of cyst nematodes, Heterodera spp. collected from five regions from Jordan (Ar-Ramtha, Madaba, Dana, Al-Karak, and Jerash) was investigated. Soil samples were collected from one representative field in each region. Morphological and morphometrical characteristics revealed that Heterodera latipons is dominated in cereal fields at Ar-Ramtha, Madaba, Dana and Al-Karak regions and Heterodera schachtii in Jerash. Cysts populations from all cereal fields had bifenestrate vulval cone and a strong underbridge. Wherever, cysts of the cabbage population had ambifenestrate vulval cone with long vulval slit. The bullae were absent in Ar-Ramtha, Madaba and Dana populations, but present in Al-Karak and Jerash. Based on 12 morphometrical characters, the first three functions in canonical discriminant analysis accounted 99.3% of the total variation. Distance from dorsal gland duct opening to stylet base, underbridge length, a = L/W (body length/midbody width) and length of hyaline tail tip had strong and significant contributions in the first function. While the second function was strongly influenced by length of hyaline tail, fenestral length, fenestral width and tail length. However, the third canonical discriminate function was found to be influenced by stylet length, fenestral length, a = L/W (body length/midbody width) and underbridge width. The graphical representation of the distribution of the samples showed that the first canonical discriminant function clearly separated H. schachtii from Jerash from other populations. Whereas, H. latipons collected from Madaba and Dana were clearly separated in the second function. The results indicated that differences at morphological and morphometrical levels revealed diverse populations of Heterodera spp. in Jordan.     

Development, adult size and fecundity of Aphidius sonchi reared in two instars of its aphid host, Hyperomyzus lactucae

Development, adult size and fecundity of the aphid parasite, Aphidius sonchi Marshall (Hymenoptera: Aphidiidae), which began their development in the first and third instar nymphs of Hyperomyzus lactucae (L.) (Homoptera: Aphididae), were studied in the laboratory. Survival and development times were almost identical in the two host instars. However, adult parasites reared in the first instar hosts were smaller and had fewer eggs at emergence than those reared from the third instar.Further analysis showed that the size of an adult parasite, irrespective of host stage at reception of the parasite egg, was largely determined by the size of the host when the parasite was in its destructive feeding phase. Furthermore, within parasites reared from each of the two host instars, the earlier emerging individuals were larger and contained more eggs on emergence. However, the relationships between size, development time and fecundity in the parasites differed significantly between cohorts reared from the two host instars.These results indicate that (1) the suitability of the aphid to the development of the parasite varies as the aphid develops, and (2) several aspects of the parasite biology must be studied to show the relative suitability of the aphids of a given instar.

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Résumé L'étude a porté sur le développement, la fécondité et la taille des adultes d'A. sonchi Marshall (Hym: Aphidiidae) qui commence son développement dans le ler et le 3ème stades larvaires d'H. lactucae (Homopt: Aphididae). La survie et la durée de développement sont presque identiques dans les hôtes des deux stades. Cependant les adultes élevés dans le ler stade de l'hôte étaient plus petits et avaient moins d'oeufs à l'émergence que ceux élevés dans le 3ème stade.Des observations ultérieures ont montré que la taille de l'adulte, indépendamment du stade de l'hôte au moment de la réception de l'oeuf, était fortement déterminée par la taille de l'hôte au moment où la larve du parasite était dans sa phase alimentaire destructrice. De plus, parmi les parasites élevés à partir des hôtes de chacun des deux stades, les premiers émergés étaient plus gros et contenaient plus d'oeufs. Cependant la relation entre taille, durée de développement et fécondité des parasites différait significativement suivant les cohortes élevées à partir des hôtes des deux stades.Ces résultats indiquent (1) que l'adéquation du puceron au développement du parasite change pendant que le puceron se développe et (2) que différents aspects de la biologie du parasite doivent être étudiés pour montrer l'adéquation relative des pucerons d'un stade donné.

     

Identification of two clip domain serine proteases involved in the pea aphid's defense against bacterial and fungal infection

Phenoloxidases (POs) are required for the pea aphid's defense against bacterial and fungal infection. Prophenoloxidases (PPOs) are proteolytically converted to its active form PO through a clip domain serine protease cascade. In this study, we identified five clip domain serine proteases in the pea aphids. The messenger RNA levels of two of them, Ap_SPLP and Ap_VP, were upregulated by Gram‐positive bacterium Staphylococcus aureus and fungus Beauveria bassiana infections. Double‐stranded RNA‐based expression knockdown of these two genes resulted in reduced PO activity of the aphid hemolymph, higher loads of S. aureus and B. bassiana in the aphids, and lower survival rates of the aphids after infections. Our data suggest that Ap_SPLP and Ap_VP are involved in PPO activation pathway in the pea aphid.     

Sieve element occlusion provides resistance against Aphis gossypii in TGR-1551 melons

Feeding behavior and plant response to feeding were studied for the aphid Aphis gossypii Glover on susceptible and resistant melons(cv.Iroquois and TGR-1551,respectively).Average phloem phase bout duration on TGR-1551 was<7% of the duration on Iroquois.Sixty-seven percent of aphids on TGR-1551 never produced a phloem phase that attained ingestion(EPG waveform E2)in contrast to only 7% of aphids on Iroquois.Average bout duration of waveform E2(scored as zero if phloem phase did not attain E2)on TGR-1551 was<3% of the duration on Iroquois.Conversely,average bout duration of EPG waveform El(sieve element salivation)was 2.8 times greater on TGR-1551 than on Iroquois.In a second experiment,liquid nitrogen was used to rapidly cryofix leaves and aphids within a few minutes after the aphids penetrated a sieve element.Phloem near the penetration site was then examined by confocal laser scanning microscopy.Ninety-six percent of penetrated sieve elements were occluded by protein in TGR-1551 in contrast to only 28% in Iroquois.Usually in TGR-1551,occlusion was also observed in nearby nonpenetrated sieve elements.Next,a calcium channel blocker,trivalent lanthanum,was used to prevent phloem occlusion in TGR-1551,and A.gossypii feeding behavior and the plants phloem response were compared between lanthanum-treated and control TGR-1551.Lanthanum treatment eliminated the sieve element protein occlusion response and the aphids readily ingested phloem sap from treated plants.This study provides strong evidence that phloem occlusion is a mechanism for resistance against A.gossypii in TGR-1551.