Improved protocols for transformation of indica rice mediated by Agrobacterium tumefaciens

A highly efficient gene transfer method mediated by Agrobacterium tumefaciens was developed for Group I indica rice, which had been quite recalcitrant in tissue culture and transformation. Freshly isolated immature embryos from plants grown in a greenhouse were inoculated with A. tumefaciens LBA4404 that harbored super-binary vector pTOK233 or pSB134, which had a hygromycin-resistance gene and a GUS gene in the T-DNA. The efficiency of gene transfer varied with the kinds of gelling agents and the basic compositions of co-cultivation media. The highest activity of GUS after co-cultivation was observed when NB medium solidified with agarose was used. For the subsequent cultures, two types of media (modified NB and CC) were chosen to recover hygromycin-resistant cells efficiently. The transformation protocol thus developed worked very well in all of the varieties tested in this study, and the transformation frequency (number of independent hygromycin-resistant and GUS-positive plants per embryo) reached more than 30% in IR8, IR24, IR26, IR36, IR54, IR64, IR72, Xin Qing Ai 1, Nan Jin 11, and Suewon 258. Most of the transformants (T₀) were normal in morphology and fertile. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the T₀ and T₁ generations. For the recovery of multiple independent transgenic events from a single immature embryo, procedures were developed to section the embryo into as many as 30 pieces after non-selective cultures following co-cultivation. Transformants were then obtained from the pieces cultured on the selective media, and, in the highest case, more than seven independent transgenic plants per original embryo (transformation frequency of 738%) were produced. Thus, the efficiency of transformation was remarkably improved.     

Feeding inhibition of Helicoverpa armigera （HiJbner） by Eucalyptus camaldulensis and Tylophora indica extracts

Crude leaf extracts of Eucalyptus camaldulensis and Tylophora indica were evaluated for their antifeedancy against Helicoverpa armigera larvae in no-choice bioassays. Cabbage leaf-disc dipped in different concentrations of leaf extracts was used for feeding fifth instars of H. armigera for 24 hours. Ethanol extracts from both the plant species exhibited significant feeding inhibition, activity. The effective concentrations of ethanol extracts that caused 50% reduction in larval feeding （EI50） were 6.9% for E. camaldulensis and 2.8% for T. indica, while these values of hexane extracts were 9.3% and 5.2%, respectively. Compared to the crude leaf extracts, crude alkaloids from T. indica and crude tannins from E. camaldulensis were more potent in reducing larval feedings.     

Differential susceptibility of subterranean termite castes to entomopathogenic nematodes

We examined the attachment and infectivity of two entomopathogenic nematode species, Steinernema carpocapsae and Heterorhabditis indica, on soldiers and workers in two subterranean termite species, Coptotermes formosanus and C. vastator. In attachment tests with S. carpocapsae, more nematodes attached to soldiers of C. formosanus and C. vastator in the absence of workers compared to soldiers that were in the presence of workers. In tests with soldiers alone, workers alone, and mixture of equal numbers of workers and soldiers, soldiers in the mixed groups had lower mortality than soldiers alone after 1 and 4 days for both termite species. Exposure of small groups of either termite species to S. carpocapsae resulted in higher mortality of soldiers after 1 and 4 days post exposure. Mortality in soldiers alone exposed to H. indica was not significant compared to mortality in mixed groups at 1 day exposure, but was significant after 4 days. In concentration-mortality tests, a significant two-way interaction existed between nematode concentration and termite caste for C. formosanus exposed to S. carpocapsae for 1 and 4 days. A significant effect of nematode concentration was found for C. vastator exposed to H. indica for 1 day. This termite species had lower mortality when exposed to H. indica after 1 and 4 days compared to C. formosanus. At 4 days post exposure to H. indica, the effects of nematode concentration and termite caste were significant. Steinernema carpocapsae caused higher mortality than H. indica, but mortality of workers was higher when exposed to H. indica. Soldiers of both species experienced rapid mortality when exposed to S. carpocapsae, whereas soldiers of C. vastator experienced lower mortality when exposed to H. indica. Thus, our results show that when soldiers alone or workers alone are exposed to the nematodes, there is a differential susceptibility of soldiers and workers to nematode infection with soldiers being more susceptible than workers. In a mixed group of soldiers and workers or workers alone, it appears that grooming behavior of the workers serves as a mechanism to reduce nematode infection of soldiers and workers. The reason for this differential response to nematode infection is that soldiers do not exhibit grooming behavior.     

Modelling nonlinear dynamics of Crassulacean acid metabolism productivity and water use for global predictions

Crassulacean acid metabolism (CAM) crops are important agricultural commodities in water‐limited environments across the globe, yet modelling of CAM productivity lacks the sophistication of widely used C3 and C4 crop models, in part due to the complex responses of the CAM cycle to environmental conditions. This work builds on recent advances in CAM modelling to provide a framework for estimating CAM biomass yield and water use efficiency from basic principles. These advances, which integrate the CAM circadian rhythm with established models of carbon fixation, stomatal conductance and the soil–plant‐atmosphere continuum, are coupled to models of light attenuation, plant respiration and biomass partitioning. Resulting biomass yield and transpiration for Opuntia ficus‐indica and Agave tequilana are validated against field data and compared with predictions of CAM productivity obtained using the empirically based environmental productivity index. By representing regulation of the circadian state as a nonlinear oscillator, the modelling approach captures the diurnal dynamics of CAM stomatal conductance, allowing the prediction of CAM transpiration and water use efficiency for the first time at the plot scale. This approach may improve estimates of CAM productivity under light‐limiting conditions when compared with previous methods.     

Bacillus thuringiensis and neem seed oil (Azadirachta indica) effects on the potato tuber moth Phthorimaea operculella zeller in the field and stores

A comparative evaluation for the efficacy of Bacillus thuringiensis and neem seed oil on Phthorimaea operculella has been carried out in the field and store. These two preparations were almost equally effective on the potato tuber moth infestation. The percentage of infestation was reduced through successive application of either preparations in the field up to harvest. No synergism was observed upon using combination of the two preparations. In the store, neem seed oil (500 ppm) was highly protective and was as effective as sevin. A combination of both neem and B.t. (Delfin) significantly protects the tubers. This suggests the possible use of either neem seed oil or B.t. in combating the insect pest in the field or during storage.     

Can the red palm mite threaten the Amazon vegetation?

The red palm mite Raoiella indica Hirst (Tenuipalpidae) was first reported in the New World in 2004, dispersing quickly and widely while adopting new plant species as hosts. Since then, it has caused severe damage in this region, especially to coconut (Cocos nucifera L.). It was first found in Brazil in 2009, in the northern Amazonian state of Roraima. In the present study, native and introduced plants were sampled between March 2010 and February 2011 in sites of the 15 Roraima municipalities, to estimate its distribution and the associated mite fauna. In addition, monthly samples were taken from a coconut plantation in Mucajaí throughout the same period, for an initial appraisal of the levels R. indica could reach. It was found in 10 municipalities, on 19 plant species of four families. Six species are reported for the first time as hosts. Among the associated predators, 89.1% were Phytoseiidae, most commonly Amblyseius largoensis (Muma), Iphiseiodes zuluagai Denmark & Muma and Euseius concordis (Chant). The highest densities of R. indica, 1.5 and 0.35 mites/cm² of leaflet (approx total of 331 and 77 mites/leaflet), were reached respectively in March 2010 and February 2011. The highest density of phytoseiids on coconut (0.009 mites/cm² or about 2 mites/leaflet) was reached in November 2010. The average densities of R. indica recorded for Roraima were comparable to those reported for countries in which the mite is reportedly economically damaging. The dispersal of R. indica through the Amazon forest may result in damage to cultivated and native palms, and plants of other families, if the projected increase in both the frequency and the severity of drought events occurs. Parts of the Amazon have undergone periods of low rainfall, a condition that appears to favour the biology of this mite. Its eventual arrival to northeastern Brazil may result in heavy economic and ecological losses.     

多级溶媒回流提取制备烟用香料酸角浸膏的研究

研究了多级溶媒提取酸角浸膏,分析了酸角浸膏的主要成分,并进行了卷烟加料试验.加料实验表明卷烟的烟气柔和细腻、香气量增加、刺激降低口感改善;低沸点部分挥发性成分种类和含量明显增加,酒石酸和还原糖含量明显较高,而蛋白质和果胶含量明显较低.     

芒果子叶切段不定根形成的影响因素分析

分别用不同成熟时间、不同取材部位、不同品种、不同大小的芒果子叶切段为外植体进行不定根的诱导,以探讨影响芒果子叶切段不定根形成能力的原因.结果表明,芒果子叶切段的生根能力随着芒果成熟度的增加而逐渐提高,花后50和60 d的2.0 cm长子叶切段都无不定根形成,从花后70 d开始有不定根形成,此时生根率为28.6%,之后其生根能力迅速提高,在花后90 d生根率达到76.7%,之后生根率稳定保持直到110 d果实成熟.成熟芒果的子叶切段长度(2.0、1.0、0.5和0.2 cm)对不定根的形成有显著影响,0.2 cm的子叶切段上无不定根形成,另外3个长度的切段都有不定根形成,且生根能力随着长度的增大而逐渐提高.取材位置(靠近或远离胚轴)则对生根影响不大,且紫花芒、红芒、青皮芒等几种常见芒果品种的不定根形成能力基本相同.     

马兰叶中总黄酮的提取

本论文以提取马兰叶中的总黄酮类化合物为目的,采用纤维素酶辅助热浸提和热浸提两种方法进行提取,利用紫外分光光度法测得马兰叶中总黄酮类化合物为分别为17.14 mg/g和14.44 mg/g,纤维素酶辅助热浸提方法的平均回收率为94.60%,相对标准偏差为5.56%。