Changes in endogenous cytokinin-like substances and growth inhibitors in freesia corms during high-temperature treatment for breaking dormancy

High summer temperatures are required to break dormancy of freesia corms and cormels. Endogenous cytokinin levels were low in corms at the lifting-date, 25 May, but increased after 3 months storage at 30°C. Cytokinin levels in cormels increased gradually during summer storage at natural room temperature, reaching a maximum on 25 August when cormels had the capacity to sprout, then decreased to low levels on 25 September. Inhibitor levels in cormels decreased gradually during storage. A zeatin riboside-like compound, water-soluble cytokinins and an abscisic acid-like compound were present in freesia corms. Dormant corms, soaked for 18 h in 1000 mg/l benzyladenine solution, showed 100% sprouting. We conclude that high temperatures cause a decrease in inhibitors and an increase in cytokinins, which together result in the breaking of the dormant state of corms and cormels of freesia.     

Effect of thin twine on gill net size-selectivity analyzed with the direct estimation method

ABSTRACT: To analyze the effect of thin twine on gill net size selectivity, a series of fishing experiments using gill nets of two twine thicknesses (no. 0.8 of 0.16 mm diameter and no. 3 of 0.28 mm diameter) and three nominal mesh sizes (41 mm, 46 mm and 51 mm) was conducted in outdoor water tanks containing a rainbow trout ( Oncorhynchus mykiss ) population of known size frequency. For nets of the same nominal mesh size, mesh openings of the no. 0.8 twine nets were approximately 2 mm larger than those of the no. 3 twine nets. The effects of both mesh opening and twine thickness are combined in the conventional analysis with the selectivity curve against fish length. Selectivity against the ratio of fish girth to mesh perimeter was calculated to avoid the direct effect of changes in mesh openings due to twine diameter at the same nominal mesh size. Results demonstrated that the thinner twine gill net was more likely to capture rainbow trout of smaller girth than the mesh perimeter. Fish with girths smaller than the mesh perimeter were often retained by the net with its body tangled by the thinner twine, possibly due to the increased flexibility of the thinner twine.     

Insecticide susceptibility of surviving Cotesia plutellae (Hym: Braconidae) and Diaeretiella rapae (M'Intosh) (Hym: Aphidiidae) as affected by sublethal insecticide dosages on host insects

The effects of sublethal dosages of insecticides applied to Plutella xylostella L. (Lepidoptera: Yponomeutidae) and Lipaphis erysimi Kaltenbach (Homoptera: Aphidiidae) on the insecticide susceptibility of the surviving endoparasitoids, Cotesia plutellae Kurdjumov (Hymenoptera: Braconidae) and Diaeretiella rapae (M'Intosh) (Hymenoptera: Aphidiidae), were studied in Shangjie, Minhou, China. The susceptibility to methamidophos and the sensitivity of acetylcholinesterase (AChE) to methamidophos and dichlorvos in the adults of host insects were substantially lower than those in the two parasitoids. The host insects were treated with sublethal dosages of methamidophos in P. xylostella and of methamidophos and avermectin in L. erysimi. The cocoon formation in the two parasitoids decreased significantly, from 35.0% (control) to 13.0% (with methamidophos treatment) for C. plutellae; from 20.6% (control) to 9.0% (with methamidophos treatment) and from 24.3% (control) to 16.7% (with avermectin treatment) for D. rapae. The susceptibility to methamidophos of the resultant emerging adults of the two parasitoids was found to be significantly lower than that of the control when the parasitoids were left in contact with the same dosages of methamidophos. The average AChE activity inhibition by methamidophos and dichlorvos in 34-60 adults of the two parasitoids that emerged from the treatments (15.1% and 31.8% respectively for C. plutellae, and 21.1% and 26.9% for D. rapae) was also significantly lower than those of the controls (55.4% and 48.3% respectively for C. plutellae, and 42.9% and 51.7% for D. rapae). The bimolecular rate constant (k(i)) values of AChE to methamidophos and dichlorvos in the adults of parasitoids without the insecticide treatment were 1.78 and 1.56 times as high as those that emerged from the host insects treated with methamidophos for C. plutellae, and 1.91 and 1.66 times as high as those in the case of D. rapae. It is suggested that there is a difference in AChE sensitivity to insecticides between the resultant emerging parasitoids with and without insecticide pretreatment. Furthermore, the introduction of the insecticides to the host insects could be an important factor in the insecticide resistance development of the endoparasitoids. The natural selectivity would favour the parasitoids that had developed an insensitivity to the insecticide(s).     

Insecticide toxicity and synergism by enzyme inhibitors in 18 species of pest insect and natural enemies in crucifer vegetable crops

The toxicities of three enzyme inhibitors and their synergistic effects on four insecticides were studied by using the dry film method on field populations of 18 species of insects collected in Jianxin and Shanjie, China, from 2003 to 2005. Meanwhile, the inhibitory effects of these enzyme inhibitors on the activities of acetylcholinesterases (AChE), carboxyesterases (CarE) and glutathione-S-transferases (GST), in vivo, were also studied. In general, triphenyl phosphate (TPP) and diethyl maleate (DEM) showed low toxicities to six herbivorous pest insects, four ladybirds and eight parasitoids. Piperonyl butoxide (PB) exhibited low toxicities to the herbivorous pest insects and ladybirds, but high toxicities to the eight parasitoids. The tolerance to the insecticides in 11 pest insects and natural enemies was mainly associated with the tolerance to PB. PB showed the highest synergism on methamidophos, fenvalerate, fipronil and avermectin in nine species of pest insects and natural enemies. In general, TPP and DEM showed significant synergisms to these four insecticides in four parasitoid species. However, in contrast to their effects on the parasitoids, the synergistic effects of TPP and DEM on the four insecticides by TPP and DEM against four pest insects and one ladybird varied depending on the insect species and enzyme inhibitor. Activity of AChE, CarE or GST could be strongly inhibited, in vivo, by PB, TPP or DEM, depending on the insect species and enzyme inhibitors. From the results obtained in this study, mixed-function oxidase (MFO) was thought to play the most critical role in insect tolerances to the tested insecticides in the field. Low competition existed in the evolution of insecticide resistance in the field populations of parasitoids, as compared with herbivorous pest insects and ladybirds. Possible causes of the high synergistic effects of PB on the four classes of insecticides, based on multiattack on the activity of CarE, GST or AChE in the insect species, are also discussed.     

Porphyran as a functional modifier of a soybean protein isolate through conjugation by the Maillard reaction

Porphyran (Por) prepared from dried nori was applied as a functional modifier of a soybean protein isolate (SPI) to conjugate with SPI from defatted soybean by the Maillard reaction (79% relative humidity and 60 degrees C for 7 days). Two kinds of partially denatured conjugate (Conj 45 and Conj 63) were obtained from the reaction product by sequential extraction at pH 4.5 and pH 6.3, and the respective yield and weight ratios of the SPI and Por moieties were 8.4% and 1:1 for Conj 45 and 11.7% and 1:0.16 for Conj 63. Conj 63 demonstrated improved solubility between pH 5.0 and pH 8.0, while Conj 45 exhibited substantially complete solubility over the pH range of 2.0-8.0. Conj 63 showed more tolerance against digestion with pancreatin than SPI, whereas this was lost after denaturation. Conj 63 and Conj 45 both showed a markedly higher emulsion activity index and emulsion stability than SPI, even at pH 3.0; in particular, Conj 45 exhibited outstanding emulsifying ability. Conj 63 had about a two-fold higher calcium-binding ability than SPI, and Conj 63 and Conj 45 did not aggregate with added Ca2+ and Mg2+. It is believed that Por could be a valuable functional modifier of SPI for providing soybean protein-based liquid foods such as beverages by conjugation through the Maillard reaction.     

Substrate specificity of carp hepatopancreatic cathepsin S for peptides and myofibrillar proteins

SUMMARY: From 250 g of carp hepatopancreas, 0.29 mg of the purified enzyme was obtained. The bond specificity of cathepsin S for α-neoendorphin and neurotensin was ⁶Arg-⁷Lys and ³Tyr-⁴Glu, respectively. The cleavage sites for insulin B-chain were estimated to be the bonds at ³Asn-⁴Gln, ⁶Leu-⁷Cys, ¹²Val-¹³Glu, ¹³Glu-¹⁴Ala, ¹⁶Tyr-¹⁷Leu, ²²Arg-²³Gly, ²⁴Phe-²⁵Phe and ²⁶Tyr-²⁷Thr. P2 position on these peptides were bulky and hydrophobic amino acid residues such as Phe or Leu, small amino acid residues such as Gly, Ala and Val were also accepted in these positions. Regarding the protein substrates, cathepsin S degraded carp α-actinin, actin, tropomyosin and troponins T and I although the proteolyzing speeds were distinct from one another.     

Detection of QTLs for white-back and basal-white grains caused by high temperature during ripening period in japonica rice

There is increasing evidence that global warming affects the development of rice. High temperatures during ripening increase the ratio of undesirable chalky grains followed by deteriorating grain appearance quality. In order to detect quantitative trait loci (QTLs) controlling the occurrence of white-back and basal-white chalky grains of brown rice, QTL analysis was performed using recombinant inbred lines derived from a cross between two strains, ‘Tsukushiroman’ (sensitive to heat stress) and ‘Chikushi 52’ (tolerant of heat stress). The F₇ and F₈ lines were exposed to heat stress during the ripening period in two locations, Fukuoka and Kagoshima, in Japan. QTLs for white-back grains and basal-white grains were detected on chromosomes 1, 3, and 8, and those for basal-white grains were detected on chromosomes 2, 3, and 12. QTLs on chromosome 8 for white-back grains were shared in the plants grown in both locations. Near-isogenic lines (NILs), which harbored a segment from ‘Chikushi 52’ on chromosome 8 with the genetic background of ‘Tsukushiroman’, showed relatively lower ratios of white-back grains than ‘Tsukushiroman’. Therefore, insertion of the ‘Chikushi 52’ genomic region of the QTL on chromosome 8 can improve the quality of rice when it is grown under heat stress conditions.     

Biochemical properties of the cell wall in the Arabidopsis mutant bor1‐1 in relation to boron nutrition

We examined concentrations of boron (B) and dimerization of rhamnogalacturonan II (RG‐II), a B‐binding polysaccharide, in the cell wall of a low‐B sensitive mutant of Arabidopsis thaliana, bor1‐1, to investigate possible effects of the bor1‐1 mutation on the biochemical form of pectins in the cell wall. In the bor1‐1 mutant, B concentrations in the cell wall from shoots were lower than those in the wild type at low B supply, whereas they were similar at sufficient B supply. The amount of B present as borate ester of the RG‐II dimer (dRG‐II‐B) in the bor1‐1 mutant was lower than that in the wild type at low B supply. In the wild type, about 90 % of RG‐II was present as dRG‐II‐B, both, at low and sufficient B supply. In the bor1‐1 mutant, about 60 % of RG‐II was in its monomeric form (mRG‐II) at low B supply, whereas more than 85 % of it was present as dRG‐II‐B at sufficient B supply. However, similar as the wild type, mRG‐II derived from the bor1‐1 mutant was able to form dRG‐II‐B in vitro in the presence of borate and lead. Sugar composition of cell wall fractions was similar in both genotypes. These results suggest that the polysaccharide composition in the cell wall was not strongly affected by the bor1‐1 mutation. The observed difference in dimerization of RG‐II at low B supply is most likely due to a reduced B concentration in the shoots of the bor1‐1 mutant.     

Reduced immunogenicity of beta-lactoglobulin by conjugation with acidic oligosaccharides

Bovine beta-lactoglobulin (beta-LG) was conjugated with the acidic oligosaccharides, alginic acid oligosaccharide (ALGO) and phosphoryl oligosaccharides (POs) by the Maillard reaction to reduce the immunogenicity of beta-LG. The molar ratios of beta-LG to ALGO and POs in the conjugates were 1:6 and 1:8. The carbohydrate-binding sites in the beta-LG-ALGO conjugate were partially identified to be (60)Lys, (77)Lys, (100)Lys, (138)Lys, and (141)Lys. The isoelectric point of each conjugate was lower than that of beta-LG. CD spectra indicated that the secondary structure of beta-LG was almost maintained after conjugation. The results of fluorescence studies indicated that the conformation around Trp had not changed in each conjugate and that the surface of each conjugate was covered with a saccharide chain. Structural analyses with monoclonal antibodies indicated that the conformation around (8)Lys-(19)Trp (beta-sheet, random coil, short helix) in the conjugates had changed, whereas the native structure was maintained around (15)Val-(29)Ile (beta-sheet) and (125)Thr-(135)Lys (alpha-helix). The beta-LG-ALGO and beta-LG-POs conjugates maintained 77 and 70% of the retinol binding activity of beta-LG. Conjugation with ALGO and POs substantially enhanced the thermal stability of beta-LG. The anti-beta-LG antibody response was markedly reduced after immunization with both conjugates in BALB/c, C57BL/6, and C3H/He mice. B cell epitopes of beta-LG and the conjugate recognized in these mice were determined with 15-mer multipin peptides, and the linear epitope profiles of the conjugates were found to be similar to those of beta-LG, whereas the antibody response to each epitope was dramatically reduced. In particular, effective reduction of the antibody response was observed in the vicinity of the carbohydrate-binding sites. Conjugation of beta-LG with these acidic oligosaccharides was effective in reducing the immunogenicity of beta-LG. The conjugates obtained in this study are edible, so they would be very useful for food application.