The morphological and physiological basis of delayed pollination overcoming pre-fertilization cross-incompatibility in Nicotiana.

Delayed pollination is widely used to overcome pre-fertilization incompatibility, but its regulatory mechanisms are unclear. When Nicotiana tabacum was cross-pollinated with pollen of N. alata, the incompatibility occurring in the basal 1/4 region of the style (pollinated at anthesis: 0-day-old pistil) was overcome by delayed pollination (of 6-day-old pistil), and the morphological changes and corresponding physiological basis are explored here. The structure and ultrastructure of the pistil were observed under fluorescence microscopy and transmission electron microscopy. Differentially expressed proteins were screened with a monoclonal antibody chip for Nicotiana, and protein expression and distribution were analysed by immunofluorescence. Cellulase and pectinase activities were tested using enzyme-linked immunosorbent assay kits. The style of Nicotiana is solid in the basal region and pollen tubes grow in the extracellular spaces (ECM) of the transmitting tissue (TTS) cells. Seven of the 22 identified proteins were cell wall-associated proteins and were expressed at higher levels during pistil senescence. Cellulase and pectinase activities increased. The TTS cells in the basal 1/4 region of the 0-day-old style were polygonal and tightly arranged, with narrow ECM, but these were oval or partially dissolved in the 6-day-old pistil, leading to wider ECM and richer secretions. The increased expression of cell wall proteins and enhanced enzyme activity during pistil senescence might partially be responsible for the cells becoming oval and the ECM enlarged, providing the morphological basis for delayed pollination overcoming the pre-fertilization incompatibility between N. tabacum and N. alata.

Safety and efficacy evaluation of laparoscopy in colorectal cancer with liver metastasis.

OBJECTIVE: To study the safety and efficacy of simultaneous completion of colorectal cancer resection and liver metastasis resection by total laparoscopy. PATIENTS AND METHODS: In the observation group, 40 patients with colorectal cancer combined with liver metastasis (CRCLM) were selected to receive total laparoscopic surgery. At the same time, 40 cases were selected for laparoscopic resection of colorectal cancer and hepatic resection as control group. RESULTS: The outcomes of the two methods in the treatment of CRCLM were compared. The results showed that the difference in surgery time between the two groups was not statistically significant (p>0.05). The blood loss, drainage tube retention time and anal exhaust recovery time in the observation group were significantly less than those in control group (p<0.05). No significant difference in completion rate was found between the two groups (p>0.05); the prevalence rate of complications in the observation group was significantly lower than that in control group (p<0.05). No significant differences in the median survival period and the survival rate at 1 year, 2 years and 3 years after surgery were found between the two groups (p>0.05). CONCLUSIONS: The outcomes of total laparoscopy in the treatment of CRCLM are not inferior to open surgery.

Changes in the Strength of Lettuce Endosperm during Germination.

The forces required to puncture intact lettuce (Lactuca sativa) seed and pericarp, endosperm and embryo were measured by the Instron Universal Testing Machine. It required about 0.6 newton to puncture the endosperm in seeds imbibed in the dark at 6, 12 and 24 hours. Endosperm of seeds imbibed in the light or in dark with gibberellic acid required about 4.2 newtons at 6 and at 12 hours and only about 0.15 newton at 24 hours. Forces required to puncture embryo at all treatments and times remained constant at about 0.3 newton. Changes in the strength of the endosperm do not appear to be related directly to protrusion of the radicle.

Differential effects of actinomycin d and cordycepin in lettuce seed germination and RNA synthesis.

Intact lettuce seed germination was inhibited by cordycepin but not by actinomycin D; however, when seeds were clipped at the cotyledonary end, actinomycin D partially inhibited germination. Uptake studies with intact seeds using (3)H-actinomycin D showed that it was unable to reach the embryo prior to radical protrusion. (3)H-Cordycepin uptake studies using intact seeds showed that cordycepin was able to reach the embryo during the first 3 hours of incubation and at subsequent times. The pericarp and endosperm offered resistance to penetration of cordycepin into the embryo. In contrast to actinomycin D, cordycepin markedly inhibited (3)H-uridine incorporation into RNA of intact seeds during the first 10 and 12 hours of incubation. About 60% of (3)H-adenosine incorporation into poly A-RNA was inhibited by cordycepin during 12 hours of incubation, whereas actinomycin D had little effect. RNA synthesis appears to be essential for seed germination.

Changes in Isoperoxidases during Cold Treatment of Dormant Pear Embryo.

The number of isoperoxidases and the intensity of certain isozymes increased with increasing periods of stratification of pear (Pyrus communis cv. Bartlett) embryos. The presence of GA(3) or kinetin during stratification enhanced the activity of certain isoperoxidases, and these enhancements were blocked in the presence of ABA which by itself had an inhibitory effect. Enhancement in isoperoxidases of pear embryos during stratification was inhibited by 6-methylpurine and cycloheximide; and in the presence of either of these two inhibitors, stratification failed to release the dormancy. Pear embryos germinated for 3 days showed changes in the pattern of isoperoxidases depending on the length of stratification.

Occurrence of some enzymes in starchy endosperm and hormonal regulation of isoperoxidase in aleurone of wheat.

Peroxidase, indoleacetic acid-oxidase, alkaline, and acid phosphatases were detected in dry starchy endosperm (minus aleurone) of wheat grain. The isoperoxidase pattern differed in different parts of the dry grain. Several new isoperoxidases were found in embryos after soaking. The intensity of isoperoxidases in aleurones was enhanced in the presence of embryo or 2 muM GA(3) after 24 hours of soaking, but decreased after 72 hours. Indoleacetic acid and kinetin had no effect on isoperoxidase of aleurone. Actinomycin D and cycloheximide had no effect on isoperoxidases of aleurones from embryonectomized or naturally occurring embryoless grains. However, these two inhibitors increased the intensity of isoperoxidases in aleurones of intact embryonated grains after soaking.

Penetration of dry seeds with chemicals applied in acetone.

Acetone is able to carry chemicals through the seed coats to reach the embryo. The amount of chemical penetrating into the seeds depends on the seed type, permeation time, and the concentration of solution. The chemicals reaching the embryo are biologically effective.

Application of chemicals in organic solvents to dry seeds.

Various chemicals were applied to dry seeds by means of organic solvents. The gibberellic acid-treated (1 mm) lettuce seeds (Lactuca sativa L.) germinated nearly 100% in the dark even after prolonged storage, and those treated with abscisic acid (1 mm or 0.5 mm) failed to germinate in the light. The seedlings emerging from morphactin-treated (1 mm) cucumber seeds (Cucumis sativus L.) exhibited profound changes in morphology. Different combinations of hormones applied to lettuce seeds caused a promotion or an inhibition of germination. Germination promotion or inhibition studies showed that the applied chemicals could be removed by washing with an organic solvent or water. Progressively larger amounts of chemicals were removed with increasing periods of washing. Thus the chemical appeared to penetrate the seed to some degree. The potential of the organic solvent method is discussed.

Extensive charging of transfer ribonucleic acid by bean leaf extracts in vitro.

1. Phenol was effectively removed from aqueous extracts of RNA by chromatography on Sephadex G-50. 2. Elution of tRNA from Sephadex G-50 columns at pH7.6 was shown to remove 91% of the endogenously bound amino acids. 3. tRNA prepared without recourse to ethanolic precipitation was capable of accepting much greater amounts of amino acids than could redissolved samples of precipitated tRNA. 4. Aminoacyl-tRNA synthetase enzymes were partially purified with calcium phosphate gel. Elution of enzymes from the gel at pH6.5 yielded a fraction having phenylalanine- and alanine-charging activity, but no aspartate-, lysine- or proline-charging activity, whereas elution at pH7.6 gave a fraction having aspartate-, lysine- and proline-charging activity but no phenylalanine- or alanine-charging activity. 5. By using partially synthetase enzymes and tRNA eluted from DEAE-Sephadex A-50 columns, 52% of the theoretical maximum of aminoacyl-tRNA synthesis was obtained in vitro.

Factors controlling aminoacyl-transfer-ribonucleic acid synthesis in vitro by a plant system.

1. Factors affecting aminoacyl-tRNA synthesis in vitro by cell-free preparations from bean leaves were investigated. 2. Evidence was obtained that optimum concentrations as well as correct ratios of Mg(2+) and ATP are required for aminoacyl-tRNA synthesis in the bean-leaf system. 3. The results indicated that pH is a controlling factor having differential effects on the formation of individual aminoacyl-tRNA species. The possible micro-regulatory function of pH in protein synthesis in vivo is discussed with special reference to alanyl-tRNA formation. 4. Very low rates of alanine-stimulated pyrophosphate exchange were observed in the absence of tRNA. This observation is discussed relative to proposals about the mechanism of aminoacyl-tRNA synthesis.

Rates of aminoacyl-transfer-ribonucleic acid synthesis in vivo and in vitro by bean leaves.

1. A procedure for measuring rates of aminoacyl-tRNA synthesis in vitro and in intact leaves is presented. 2. Leaf discs showed rates close to those of intact leaves. 3. Cell-free preparations showed similar rates when assayed by pyrophosphate exchange, but actual aminoacyl-tRNA formation rates appeared to be much lower. Evidence is presented that dilution of supplied labelled amino acids was a major factor causing the low apparent rates. 4. Attempts to strip endogenous amino acids from plant tRNA resulted in low acceptor capability of the tRNA.