l-tryptophan-assisted PGPR-mediated induction of drought tolerance in maize (Zea mays L.)

Drought is an important abiotic stress that limits the plant growth and productivity. Present investigation was aimed that plant growth-promoting rhizobacteria (PGPR) isolated from moisture-stressed area impart drought tolerance in plants and tryptophan may improve their efficiency. Pseudomonas sp. (1), Bacillus cereus and Bacillus pumilus (B. pumilus) were isolated from maize rhizosphere grown in irrigated fields, semi-arid region and arid region, respectively. Proteus sp. and Pseudomonas sp. (2) were isolated from rice rhizosphere grown in irrigated fields and raised bed. B. pumilus produced 5× more abscisic acid (ABA) in culture media than Pseudomonas sp. (1) by the addition of l-tryptophan. These inoculants also modulated the phytohormone content of maize leaves in a pot experiment. Higher ABA was produced by the application of B. pumilus and Pseudomonas sp. (2), while indole 3-acetic acid and gibberellic acid were found higher in Pseudomonas sp. (1) and Proteus sp. treated plants. Addition of l-tryptophan increased the concentration of all phytohormones in soil and leaves of maize. Maximum increase in relative water content, osmotic potential, protein content and photosynthetic pigments was recorded in B. pumilus treated maize plants. Under irrigated condition, response of Pseudomonas sp. co-inoculated with B. pumilus from arid field superseded while under drought stress the effect of later predominated. Bacillus pumilus can be used in the formulation of biofertilizer to alleviate drought stress in arid and semi-arid regions.     

In vitro pollination of maize (Zea mays L.) — Proof of double fertilization

Isolated ovules from the maize homozygous recessive brown midrib (bm₃) were in vitro pollinated by pollen carrying the dominant allele — and the purple embryo marker (PEM). The colour characters of the embryos and kernels corresponded to the results of control pollination and confirmed the process of double fertilization. The question whether the method is useful for obtaining haploids is discussed.     

Plastid targeting of E. coli β-glucuronidase and ADP-glucose pyrophosphorylase in maize (Zea mays L.) cells

Summary Dicot and monocot chloroplast targeting peptides (CTPs) were evaluated for their effect on targeting, processing, and expression of two reporter proteins in maize cells. When tested transiently in maize leaf protoplasts, the maize ribulose bisphosphate carboxylase small subunit CTP required the inclusion of the amino terminus of mature small subunit protein to target -glucuronidase (GUS) to the plastid. To remove this amino terminal extension from GUS after import and processing, a repeat of the native processing site was inserted between the native mature protein and the reporter protein. This repeat processing site was used with less efficiency than the native site. Parallel constructs using the Arabidopsis thaliana small subunit and maize granule-bound starch synthase CTPs also localized GUS, but varied in repeat site use and GUS expression levels. Data from the CTP fusions with GUS were generally confirmed with fusions to an allosteric variant of E. coli ADP-glucose pyrophosphorylase. Plastid targeting of this enzyme was required for starch enhancement of transgenic BMS cells.Abbreviations BMS maize Black Mexican Sweet suspension culture cells - CTP chloroplast targeting peptide - glgC16 an allosteric variant of E. coli ADP-glucose pyrophosphorylase - GUS -glucuronidase - LUX luciferase - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - SSU small subunit of ribulose bisphosphate carboxylase     

Genetic control and racial variation of β-glucosidase isozymes in maize (Zea mays L.)

-Glucosidase (-D-glucoside glucohydrolase, E.C. 3.2.1.21, -Glu) isozyme variants were studied in a large number of inbred lines, crosses, and races of maize (Zea mays L.). The pattern of Mendelian inheritance demonstrated for -GLU variants indicated that they are under nuclear gene control. Twenty-two allelic forms at a single locus were identified in the materials studied by starch gel electrophoresis. Genetic data indicate that -GLU in maize is functionally a dimer. Variation of -GLU isozymes in 51 racial collections of maize from Mexico showed little correlation with morphological or geographical data. In 39 collections from Central America, variation patterns appeared to have some association with altitude.This work was supported in part by NIH Research Grant GM 11546.Paper No. 5040 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, North Carolina.     

Are cytological parameters of maize landraces (Zea mays ssp. mays) adapted along an altitudinal cline?

Journal of Plant Research - The Northwestern Argentina (NWA) highland region is one of the southernmost areas of native maize cultivation. We studied variations of different cytological parameters,...     

Intracellular immunolocalization of adenosine 5′-diphosphoglucose pyrophosphorylase in developing endosperm cells of maize (Zea mays L.)

We present immuno-electron microscopic evidence to show that ADPglucose pyrophosphorylase (AGP, EC 2.7.7.27) encoded by the Sh2 (shrunken 2) and the Bt2 (brittle 2) genes is localized to amyloplasts in developing endosperm of maize. The three AGP antibodies, including the two maize antisera, each raised against the Sh2encoded large or the Bt2-encoded small subunit and the spinach leaf protein, showed strong immuno-gold signals on developing starch grains in amyloplasts. The maize antibodies, but not the spinach, detected additional cross-reactivity sites to endosperm cell wall. Similar endosperm sections of the sh2-null mutant, lacking the Sh2-encoded subunit, yielded a drastic reduction in immuno signal on both starch grains and cell wall with the Sh2 anti-serum. However, the Bt2 and the spinach antisera showed no detectable difference between the sh2-null and the wild-type genotypes, except that the spinach antisera showed no reactivity to the cell wall in either of the two genotypes. Because the Bt2 epitope was readily detectable on the sh2-null starch grains, we suggest that the Bt2 subunit of this heteromeric enzyme is able to target itself to the organelle. The amyloplastic localization of the AGP protein in our studies is given additional significance by recent molecular data which indicate that full-length cDNA clones of the Sh2 and Bt2 genes show no cleavable transit-peptide signal sequence in their deduced aminoacid sequences. The observed disparity between the molecular and immunocytochemical data described here is discussed in the context of other proteins engaged in intracellular translocation with and without the known signal sequences.     

Down-Regulation of ZmEXPB6 (Zea mays β-Expansin 6) Protein Is Correlated with Salt-mediated Growth Reduction in the Leaves of Z. mays L.

The salt-sensitive crop Zea mays L. shows a rapid leaf growth reduction upon NaCl stress. There is increasing evidence that salinity impairs the ability of the cell walls to expand, ultimately inhibiting growth. Wall-loosening is a prerequisite for cell wall expansion, a process that is under the control of cell wall-located expansin proteins. In this study the abundance of those proteins was analyzed against salt stress using gel-based two-dimensional proteomics and two-dimensional Western blotting. Results show that ZmEXPB6 (Z. mays β-expansin 6) protein is lacking in growth-inhibited leaves of salt-stressed maize. Of note, the exogenous application of heterologously expressed and metal-chelate-affinity chromatography-purified ZmEXPB6 on growth-reduced leaves that lack native ZmEXPB6 under NaCl stress partially restored leaf growth. In vitro assays on frozen-thawed leaf sections revealed that recombinant ZmEXPB6 acts on the capacity of the walls to extend. Our results identify expansins as a factor that partially restores leaf growth of maize in saline environments.     

Optimisation of DNA transfer and transientβ-glucuronidase expression in electroporated maize (Zea mays L.) microspores

Summary The ability to deliver free DNA into microspores of a highly androgenic hybrid of maize was assessed by electroporation, using a square wave pulse discharge apparatus. The electroporation medium was chosen according to its ability to maintain a high level of regeneration. Nuclease activities were analyzed and were inhibited by the addition of 100 mM KNO₃ and MgSO₄ in the electroporation medium. Seven expression vectors withUid A as the reporter gene under the control of cauliflower mosaic virus 35S, Lat 52-7, Zmg 13, Emu, Ubiq-1, Al, or Actl promoters were tested in relation to the level of ß-glucuronidase expression in maize microspores. The highest level of expression was obtained when theUid A gene was driven by the Actl promoter. Therefore, this vector was further used to define optimal conditions leading to highest levels of ß-glucuronidase expression. The parameters determined in this study could provide an ideal starting point for the obtention of transgenic maize plants from electroporated microspores.Abbreviations DH diplohaploid - PEG polyethylene glycol - GUS ß-glucuronidase - EDTA Ethylene-diaminetetra Acetic acid - CaMV Cauliflower mosaic virus     

Measurement of N2 fixation in maize (Zea mays L.)—ricebean (Vigna umbellata [Thunb.] Ohwi and Ohashi) intercrops

The yield of N in maize (Zea mays L.) and ricebean (Vigna umbellata [Thumb.] Ohwi and Ohashi) were compared on a Tropoqualf soil in North Thailand in 1984 and 1985. Both species were grown in field plots in monoculture or as intercrops at a constant planting density equivalent to 8 maize or 16 ricebean plants per m². The contribution of symbiotic N₂ fixation to ricebean growth was estimated from measurements of the natural abundance of¹⁵N (δ¹⁵N) in shoot nitrogen and from analysis of ureides in xylem sap vacuumextracted from detached stems. The natural abundance of¹⁵N in the intercropped ricebean was found to be considerably less than that in monoculture in both growing seasons. Using maize and a weed (Ageratum conyzoides L.) as non-fixing¹⁵N reference plants the proportions (P ¹⁵N) of ricebean shoot N derived from N₂ fixation ranged from 0.27 to 0.36 in monoculture ricebean up to 0.86 when grown in a 75% maize: 25% ricebean intercrop. When glasshouse-derived calibration curves were used to calculate plant proportional N₂ fixation (Pur) from the relative ureide contents of field collected xylem exudates, the contribution of N₂ fixation to ricebean N yields throughout the 1985 growing season were greater in intercrop than in monocrop even at the lowest maize:legume ratio (25∶75). Seasonal patterns of sap ureide abundance indicated that N₂ fixation was greatest at the time of ricebean podset. The averagePur andP ¹⁵N in ricebean during the first 90 days of growth showed identical rankings of monocrop and intercrop treatments in terms of N₂ fixation, although the two sets ofP values were different. Nonetheless, seasonal estimates of N₂ fixation during the entire 147 days of legume growth determined from ureide analyses indicated that equivalent amounts of N could be fixed by ricebean in a 75∶25 intercrop and in monoculture despite the former being planted at one-quarter the density.     

(—) S-adenosyl-L-methionine-magnesium Protoporphyrin Methyltransferase, an Enzyme in the Biosynthetic Pathway of Chlorophyll in Zea mays

The enzyme (—) S-adenosyl-L-methionine-magnesium protoporphyrin methyltransferase, which catalyzes the transfer of the methyl group from (—) S-adenosyl-L-methionine to magnesium protoporphyrin to form magnesium protoporphyrin monomethyl ester, has been detected in chloroplasts isolated from Zea mays.

Zinc protoporphyrin and free protoporphyrin also act as substrates in the system, although neither one is as active as magnesium protoporphyrin.

The following scheme of chlorophyll synthesis in higher plants is proposed: δ-aminolevulinic acid → → → protoporphyrin → magnesium protoporphyrin → magnesium protoporphyrin monomethyl ester → → → chlorophyll a.

     

Starch Mobilization in Ultradried Seed of Maize （Zea mays L.） During Germination

The effects of ultradry storage on the starch mobilization in maize (Zea mays L.) seed after aging were investigated. The results indicated that there were no significant differences in the content of ATP, starch, and soluble sugar, as well as the activity of amylase, between ultradried seeds and seeds stored at -20℃ during germination. These results were consistent with the higher level of vigor of the ultradried seed. Sieve tube introduction of a fluorescence dye (carboxyl fluoresceindiacetate) and laser confocal microscopy were used to study the development of plasmodesmata in the ultradried seeds. The results indicated that plasmodesmata developed well in ultradried seeds. Fluorescence analysis also showed that the fluorescence intensity in the radicle of ultradried seeds was stronger than that in seeds with a higher moisture content. This suggests that ultradry treatment has no adverse effects on the seeds. After seed imbibition, cell orgaelles could be resumed. It is concluded that ultradry seed storage is beneficial for maintaining seed vigor and that starchy mobilization proceeds regularly during germination.     

Interaction between NO 3 − and abscisic acid in the regulation of lateral root elongation in Zea mays L.

Lateral root (LR) elongation rate of 7–8-day maize seedlings depends on the availability of NO ₃ ^? , NO ₂ ^? , and abscisic acid (ABA) in an environment. Four-hour exposure to 0.01–1.5 mM NO ₂ ^? increases the relative LR elongation rate; in the case of NO ₂ ^? , the stimulation occurs only at an NO ₂ ^? concentration equal to 0.01 mM. Exogenous ABA (10^?6 M) inhibits the LR elongation process. In the case of a combined influence of NO ₃ ^? and ABA or NO ₂ ^? and ABA, the character of the response elongation reaction is different. The NO role in the regulation of LR elongation is discussed.     

Enzyme des Stärkeumsatzes in den Wurzelhaubenzellen von Zea mays L.

Summary In connection with the problem of the well-known stability of statolith starch, some enzymes of starch metabolism have been investigated qualitatively in the root cap cells of Zea mays L. No activity of granule-bound UDPG- and ADPG-transglucosylase (EC 2.4.1.21) could be found. In the soluble enzyme fraction of the root cap cells, on the other hand, activities of phosphorylase (EC 2.4.1.1), sucrose synthetase (EC 2.4.1.13), UDPG-pyrophosphorylase (EC 2.7.7.9), -Amylase (EC 3.2.11), Maltase (EC 3.2.1.20), and D-enzyme (EC 2.4.1.25) were clearly shown to be present. However, no measurable activities of ADPG-pyrophosphorylase, sucrose-6-phosphate-synthetase (EC 2.4.1.14) and UDPG-dehydrogenase (EC 1.1.1.22) could be found. It is concluded that the stability of statolith starch in the root cap cells is not caused by the lack of enzymes of starch metabolism, but perhaps by a dynamic equilibrium between the degradation and the synthesis of starch. The later could proceed by the activity of phosphorylase working in the direction of starch synthesis because of removal of the inorganic phosphate by phosphorylating mitochondria accumulating in the neighbourhood of the statolith amyloplasts.     

Further characterization of AFLP® data as a tool in genetic diversity assessments among maize (Zea mays L.) inbred lines

AFLP® markers generated by CNG methylation sensitive (PstI/MseI) and CNG methylation insensitive (EcoRI/MseI) enzyme combinations and AFLP markers collected from hypomethylated (PstI/MseI) and hypermethylated (^m PstI/MseI) regions were compared for their polymorphism information content, sampling variance and patterns of genetic diversity in a representative sample of 33 inbred lines of maize (Zea mays L.). We demonstrate that the mean polymorphism information content generated by sets of PstI/MseI and ^m PstI/MseI markers (0.38) is significantly higher than by sets ofEcoRI/MseI markers (0.33). Also the sampling variance highlighted the distinctive nature of the ^(m) PstI/MseI markers: to achieve a mean standard deviation of 5% in the estimation of genetic distance among the 33 inbreds, the PstI/MseI and ^m PstI/MseI marker sets (135 and 129 markers, respectively) are clearly smaller than the EcoRI/MseI marker set (173 markers). A further minimizing of the sampling variance of AFLP data in the estimation of genetic similarities was obtained by reducing marker information redundancy by selecting markers evenly distributed over each chromosome: a set of only 106 AFLP markers, sampled conditionally on their genetic map position, was required for a mean standard deviation of 5% in the estimation of genetic distance among the 33 inbreds.     

Regulation of Glutamate Dehydrogenase Activity in Maize Leaves (Zea mays L.) with Change in the Light Сonditions

Biology Bulletin - The molecular and biochemical mechanisms of regulation of the activity of glutamate dehydrogenase (GDH) in maize leaves were studied when the light regime was changed. The...     

Effects of γ-irradiation on elongation and indole-3-acetic acid level of maize (Zea mays) coleoptiles

The effects of γ-irradiation on elongation and the level of indole-3-acetic acid (IAA) of maize (Zea mays) coleoptiles were investigated. When 3-day-old seedlings of maize were exposed to γ-radiation lower than 1 kGy, a temporal retardation of coleoptile elongation was induced. This retardation was at least partly ascribed to a temporal decrease in the amount of free IAA in coleoptile tips on the basis of the following facts: (1) the reactivity to IAA of the elongating coleoptile cells was not altered by irradiation; (2) endogenous IAA level in the tip of irradiated coleoptiles was at first unchanged, but then declined before returning to nearly the same level as that of the non-irradiated control; and (3) the amount of IAA that diffused from coleoptile tip sections showed a similar pattern to that of endogenous IAA. The rate of conversion between free and conjugated IAA was not significantly affected by irradiation. These results suggest that a temporal inhibition of maize coleoptile elongation induced by γ-irradiation can be ascribed to the reduction of endogenous IAA level in the coleoptile tip, and this may originate from the modulation in the rate of IAA biosynthesis or catabolism.