大豆体细胞胚胎发生与农杆菌介导的遗传转化

以55个大豆基因型未成熟子叶为外植体,用高浓度2,4-D诱导大豆体细胞胚胎发生与植株再生,并对生产上种植面积大、体细胞胚胎发生率高的大豆基因型用农杆菌介导法进行遗传转化。结果表明,东北地区主栽的大豆基因型中有14个基因型体细胞胚胎发生率超过40%。用含有pGBI121S4ABC质粒的LBA4404农杆菌侵染5个东北地区主栽大豆基因型的2147个未成熟子叶,经卡那霉素抗性筛选得到12株PCR阳性植株。 Abstract：Somatic embryogenesis was induced and the regenerated plants were obtained by higher concentrations of auxins with immature cotyledon of 55 genotypes in soybean. Bivalent insect resistant genes were transformed into immature cotyledon of soybean which have high frequency of somatic embryogenesis via Agrobacterium-mediated. The results showed that 14 genotypes possessed high frequency of somatic embryogenesis (more than 40%) among soybean genotypes from Northeast area. 2147 immature cotyledons of 5 different soybean genotypes cultured in Northeast area were inoculated with LBA4404 (including pGBI121S4ABC plasmid). 12 regenerated plants selected by Kanamicy gave positive PCR reaction.     

Effect of boundary layer conductance on the response of stomata to humidity

Abstract. Leaf conductance responses to leaf to air water vapour partial pressure difference (VPD) have been measured at air speeds of 0.5 and 3.0 ms⁻¹ in single attached leaves of three species in order to test the hypothesis that leaf conductance response to VPD is controlled by evaporation from the outer surface of the epidermis, rather than by evaporation through stomata. Total conductance decreased linearly with increassing VPD at both air speeds, but was decreased 1.6 3.0 times as much as by a given incrase in VPD at high than at low air speed. depending on species. In all species the relationship between leaf conductance and the gradient for evaporation from the epidermis was the same at both values of boundary layer conductance, supporting the hypothesis that direct epidermal evaporation controls stomatal guard cell behaviour in responses of stomata to VPD in these species.     

Isolation and Partial Characterization of the Major Seed Protein from Nicotiana tabacum,and Accumulation during Development

During the seed development of Nicotiana tabacum, appreciable accumulation of the soluble protein fraction started to occur at around the 6th day after anthesis and finally reached 12% on the basis of dry weight when seed maturation was accomplished. In the soluble fraction of mature seeds, four protein fractions were observed on analytical ultracentrifugation, and the protein having a sedimentation coefficient of 11.7S was the major one. The 11.7S protein was isolated and SDS-polyacrylamide gel electrophoresis indicated that the protein consisted of at least five subunits with molecular weights of 49,000, 31,000, 29,000, 21,000 and 19,000. The 11.7S protein was rich in glutamic acid or glutamine and arginine, and the presence of carbohydrate was confirmed.

During development, all of the five subunits started to appear during the period between the 12th and 15th day after anthesis.     

Synthesis and degradation of the major allergens in developing and germinating soybean seed

Gly m Bd 28K,Gly m Bd 30K and Gly m Bd 60K are the major soybean(Glycine max(L.)Merr.)allergens limiting the consumption of a good protein source for sensitive individuals.However,little is known about their temporal-spatial expression during seed development and upon germination.The present data shows that soy allergens accumulated in both the embryonic axes and cotyledon,but expression patterns differed depending on the specific allergen.Allergens accumulated sooner and to a greater level in cotyledons than in embryonic axes.Gly m Bd 28 began at 14 d after flowering,7 to 14 d earlier than Gly m Bd 30K and Gly m Bd 60K.Comparatively,their degradation was faster and more profound in embryonic axes than in cotyledons.Gly m Bd 60K began to decline at 36 h after imbibition and remained detectable up to 108 h in cotyledons.In contrast,the Glym Bd 60K protein was reduced at 24 h,and eventually disappeared at 96 h.In cotyledons Gly m Bd 28K first declined at 24 h,then increased from 36 h to 48 h,followed by its large reduction at 72 h after seed germination.These findings provide useful information on soy allergen biosynthesis and will help move forward towards developing a hypoallergenic soybean for safer food.     

Abscisic acid mimics effects of dehydration on area expansion and photosynthetic partitioning in young soybean leaves

Abstract. Leaf area expansion, photosynthetic carbon dioxide uptake and leaf dry mass accumulation were compared for expanding leaves of well-watered soybean ( Glycine max [L.] Merr.) plants, mildly dehydrated plants and well-watered plants treated with abscisic acid (ABA). Both ABA treatment and dehydration reduced area expansion in the light and over a 24 h period without decreasing the photosynthetic rates of expanding leaves. Dry mass accumulation during the light was less in ABA-treated and water-stressed leaves than in control leaves, with no differences among treatments in leaf mass per unit of area. ABA treatment and water stress both increased export of carbon from expanding leaves in the light. ABA applied near the end of the light period also increased export of carbon during the following dark period. However, it is unlikely that decreased availability of photosynthate caused slow expansion in the ABA and dehydration treatments, because expansion rates were not slowed in plants kept in dim light, even though photosynthetic rates and dry mass accumulation rates were greatly reduced. The data suggest that ABA may mediate the effects of mild dehydration on leaf area expansion and partitioning of photosynthate.     

Assessment of potential sources of error in nitrogen fixation measurements by the nitrogen-15 isotope dilution technique

Although the use of ¹⁵N fertilizers to measure nitrogen (N₂) fixed in crops has increased substantially in recent years, some methodological uncertainties still remain unresolved. The results obtained from a greenhouse study of soybean [Glycine max. (L.) Merrill] inoculated by six different methods have been examined for potential errors arising from incorporating ¹⁵N labelled fertilizer into soil to estimate N₂ fixed in pods or shoots or the whole plant at three growth stages (50% flowering, pod-initiation and physiological maturity) using as reference crops, an uninoculated soybean cultivar and a non-nodulating soybean isoline. At the first harvest when N₂ fixed was very low, the estimates of N₂ fixed by the two reference crops did not match. At this stage the uninoculated soybean estimated about four times as much N₂ fixed in the symbiotic soybean as that measured using the non-nodulating soybean. For the second and third harvests, there were substantial increases in N₂ fixed, and both the non-nodulating and uninoculated soybean were equally suitable as reference crops for assessing N₂ fixed in the symbiotic soybean. These results indicate how critical and difficult the choice of the reference crop could be at early harvests, or when N₂ fixed is low. Even though there were significant differences in ¹⁵N enrichments in different organs (generally nodules < pods < roots < shoots), the estimates of N₂ fixed in soybean plants obtained by excluding roots and nodules did not differ much from those based on the whole plant. Of the above-ground organs, % N₂ fixed in pods (containing seeds) was closest to that of the whole plant (similar at P<0.05 at physiological maturity). However, the total N₂ fixed in pods or shoots was substantially lower than that fixed by the whole plant (P<0.05), although that for the pods and enclosed seeds once again was closer to N₂ fixed in the whole plant than that in the shoots.     

Oxygen limitation of N2 fixation in stem-girdled and nitrate-treated soybean

The effects of increasing rhizosphere pO₂on nitrogenase activity and nodule resistance to O₂diffusion were investigated in soybean plants [Glycine max (L.) Merr. cv. Harosoy 63] in which nitrogenase (EC 1.7.99.2) activities were inhibited by (a) removal of the phloem tissue at the base of the stem (stem girdling), (b) exposure of roots to 10 mM NO₃over 5 days (NO₃-treated), or (c) partial inactivation of nitrogenase activity by an exposure of nodulated roots to 100 kPa O₂(O₂-inhibitcd). In control plants and in plants which had been treated with 100 kPa O₂, increasing rhizosphere O₂concentrations in 10 kPa increments from 20 to 70 kPa did not alter the steady-state nitrogenase activity. In contrast, in plants in which nitrogenase activities were depressed by stem girdling or by exposure to NO₃, increasing rhizosphere pO₂resulted in a recovery of 57 or 67%, respectively, of the initial, depressed rates of nitrogenase activity. This suggests that the nitrogenase activity of stem-girdled and NO₃-treated soybeans was O₂-limited. For each treatment, theoretical resistance values for O₂diffusion into nodules were estimated from measured rates of CO₂exchange, assuming a respiratory quotient of 1.1 and 0 kPa of O₂in the infected cells. At an external partial pressure of 20 kPa O₂, the stem-girdled and NO₃^--treated plants displayed resistance values which were 4 to 8.6 times higher than those in the nodules of the control plants. In control and O₂-inhibited plants, increases in pO₂from 20 to 70 kPa in 10 kPa increments resulted in a 2.5- to 3.9-fold increase in diffusion resistance to O₂, and had little effect on either respiration or nitrogenase activity. In contrast, in stem-girdled and NO₃^--treated plants, increases in external pO₂had little effect on diffusion resistance to O₂, but resulted in a 2.3- to 3.2-fold increase in nodule respiration and nitrogenase activity. These results are consistent with stem-girdling and NO₃^--inhibition treatments limiting phloem supply to nodules causing an increase in diffusion resistance to O₂at 20 kPa and an apparent insensitivity of diffusion resistance to increases in external pO₂.     

Evidence supporting a non-phloem source of water for export of solutes in the xylem of soybean root nodules

The vascular anatomy of soybean nodules [Glycine max (L.) Merr.] suggests that export of solutes in the xylem should be dependent on influx of water in the phloem. However, after severing of stem xylem and phloem by shoot decapitation, export of ureides from nodules continued at an approximately linear rate for 5h. This result was obtained with decapitated roots remaining in the sand medium, but when roots were disturbed by removal from the rooting medium prior to shoot decapitation, export of ureides from nodules was greatly reduced. Stem exudate could not be collected from disturbed roots, indicating that flow in the root xylem had ceased. Thus, ureide export from nodules appeared to be dependent on a continuation of flow in the root xylem. When seedlings were fed a mixture of ³H₂O and ¹⁴C-inulin for periods of 14–21 min, nodules had higher ³H/¹⁴C ratios than roots from which they were detached. The combined results are not consistent with the proposal that export of nitrogenous compounds from nodules is dependent on import of water via the phloem. The results do support the view that a portion of the water required for xylem export from soybean nodules is supplied via a symplastic route from root cortex to nodule cortex to the nodule vascular apoplast.