Endophytic microbes: biodiversity, plant growth-promoting mechanisms and potential applications for agricultural sustainability.

Endophytic microbes are known to live asymptomatically inside their host throughout different stages of their life cycle and play crucial roles in the growth, development, fitness, and diversification of plants. The plant-endophyte association ranges from mutualism to pathogenicity. These microbes help the host to combat a diverse array of biotic and abiotic stressful conditions. Endophytic microbes play a major role in the growth promotion of their host by solubilizing of macronutrients such as phosphorous, potassium, and zinc; fixing of atmospheric nitrogen, synthesizing of phytohormones, siderophores, hydrogen cyanide, ammonia, and act as a biocontrol agent against wide array of phytopathogens. Endophytic microbes are beneficial to plants by directly promoting their growth or indirectly by inhibiting the growth of phytopathogens. Over a long period of co-evolution, endophytic microbes have attained the mechanism of synthesis of various hydrolytic enzymes such as pectinase, xylanases, cellulase, and proteinase which help in the penetration of endophytic microbes into tissues of plants. The effective usage of endophytic microbes in the form of bioinoculants reduce the usage of chemical fertilizers. Endophytic microbes belong to different phyla such as Actinobacteria, Acidobacteria, Bacteroidetes, Deinococcus-thermus, Firmicutes, Proteobacteria, and Verrucomicrobia. The most predominant and studied endophytic bacteria belonged to Proteobacteria followed by Firmicutes and then by Actinobacteria. The most dominant among reported genera in most of the leguminous and non-leguminous plants are Bacillus, Pseudomonas, Fusarium, Burkholderia, Rhizobium, and Klebsiella. In future, endophytic microbes have a wide range of potential for maintaining health of plant as well as environmental conditions for agricultural sustainability. The present review is focused on endophytic microbes, their diversity in leguminous as well as non-leguminous crops, biotechnological applications, and ability to promote the growth of plant for agro-environmental sustainability.

Consensus genomic regions associated with grain protein content in hexaploid and tetraploid wheat.

A meta-analysis of QTLs associated with grain protein content (GPC) was conducted in hexaploid and tetraploid wheat to identify robust and stable meta-QTLs (MQTLs). For this purpose, as many as 459 GPC-related QTLs retrieved from 48 linkage-based QTL mapping studies were projected onto the newly developed wheat consensus map. The analysis resulted in the prediction of 57 MQTLs and 7 QTL hotspots located on all wheat chromosomes (except chromosomes 1D and 4D) and the average confidence interval reduced 2.71-fold in the MQTLs and QTL hotspots compared to the initial QTLs. The physical regions occupied by the MQTLs ranged from 140 bp to 224.02 Mb with an average of 15.2 Mb, whereas the physical regions occupied by QTL hotspots ranged from 1.81 Mb to 36.03 Mb with a mean of 8.82 Mb. Nineteen MQTLs and two QTL hotspots were also found to be co-localized with 45 significant SNPs identified in 16 previously published genome-wide association studies in wheat. Candidate gene (CG) investigation within some selected MQTLs led to the identification of 705 gene models which also included 96 high-confidence CGs showing significant expressions in different grain-related tissues and having probable roles in GPC regulation. These significantly expressed CGs mainly involved the genes/gene families encoding for the following proteins: aminotransferases, early nodulin 93, glutamine synthetases, invertase/pectin methylesterase inhibitors, protein BIG GRAIN 1-like, cytochrome P450, glycosyl transferases, hexokinases, small GTPases, UDP-glucuronosyl/UDP-glucosyltransferases, and EamA, SANT/Myb, GNAT, thioredoxin, phytocyanin, and homeobox domains containing proteins. Further, eight genes including GPC-B1, Glu-B1-1b, Glu-1By9, TaBiP1, GSr, TaNAC019-A, TaNAC019-D, and bZIP-TF SPA already known to be associated with GPC were also detected within some of the MQTL regions confirming the efficacy of MQTLs predicted during the current study.

Effect of soaking and germination treatments on nutritional, anti-nutritional, and bioactive properties of amaranth (Amaranthus hypochondriacus L.), quinoa (Chenopodium quinoa L.), and buckwheat (Fagopyrum esculentum L.).

Pseudocereals have attracted the attention of nutritionists and food technologists due to their high nutritional value. In addition to their richness in nutritional and bioactive components, these are deficient in gluten and can serve as valuable food for persons suffering from gluten allergies. Processing treatments are considered an effective way to enhance the quality of food grains. Soaking and germination are traditional and most effective treatments for enhancing the nutritional and bioactive potential as well as reducing the anti-nutritional components in food grains. This study reflects the effect of soaking and germination treatments on nutritional, bioactive, and anti-nutritional characteristics of pseudocereals. There was a significant (p ≤ 0.05) increase in nutritional and bioactive components such as crude fiber, crude protein, phenolic components, antioxidant activity, and mineral content but reduced the anti-nutrients such as tannin and phytic acid. In amaranth, there was a significant increase (p ≤ 0.05) of 7.01, 74.67, 126.62, and 87.47% in crude protein, crude fiber, phenolic content, and antioxidant activity but significant (p ≤ 0.05) reduction of 32.30% and 29.57% in tannin and phytic acid contents, respectively. Similar changes in values of crude proteins, crude fiber, phenolic content, and antioxidant activity were observed in buckwheat and quinoa. While the anti-nutritional components such as tannin and phytic acid decreased by 59.91 and 17.42%, in buckwheat and 27.08% and 47.57%, in quinoa, respectively. Therefore, soaking and germination proved to be excellent techniques to minimize the anti-nutritional component and enhance the nutritional, bioactive, and antioxidant potential of these underutilized grains.

A candidate gene OsAPC6 of anaphase-promoting complex of rice identified through T-DNA insertion.

A dwarf mutant (Oryza sativa anaphase-promoting complex 6 (OsAPC6)) of rice cultivar Basmati 370 with 50% reduced plant height as compared to the wild type was isolated by Agrobacterium tumefaciens-mediated transformation using Hm(R) Ds cassette. This mutant was found to be insensitive to exogenous gibberellic acid (GA(3)) application. Homozygous mutant plants showed incomplete penetrance and variable expressivity for plant height and pleiotropic effects including gibberellic acid insensitivity, reduced seed size, panicle length, and female fertility. Single copy insertion of T-DNA and its association with OsAPC6 was confirmed by Southern hybridization, germination on hygromycin, and 3:1 segregation of HPT gene in F(2) from OsAPC6 x Basmati 370 cross. The T-DNA flanking region sequenced through thermal asymmetric interlaced polymerase chain reaction showed a single hit on chromosome 3 of japonica rice cultivar Nipponbare in the second exonic region of a gene which encodes for sixth subunit of anaphase-promoting complex/cyclosome. The candidate gene of 8.6-kb length encodes a 728-amino acid protein containing a conserved tetratricopeptide repeat (TPR) domain and has only a paralog, isopenicillin N-synthase family protein on the same chromosome without the TPR domain. There was no expression of the gene in the mutant while in Basmati 370, it was equal in both roots and shoots. The knockout mutant OsAPC6 interferes with the gibberellic acid signaling pathway leading to reduced height and cell size probably through ubiquitin-mediated proteolysis. Further functional validation of the gene through RNAi is in progress.

Characteristics of an Acidic Phytase from Aspergillus aculeatus APF1 for Dephytinization of Biofortified Wheat Genotypes.

Phytases are the special class of enzymes which have excellent application potential for enhancing the quality of food by decreasing its inherent anti-nutrient components. In current study, a protease-resistant, acidic phytase from Aspergillus aculeatus APF1 was partially purified by ammonium sulfate fractionation followed by chromatography techniques. The molecular weight of partially purified phytase was in range of 25-35 kDa. The purified APF1 phytase was biochemically characterized and found catalytically active at pH 3.0 and 50 °C. The Km and Vmax values of APF1 phytase for calcium phytate were 3.21 mM and 3.78 U/mg protein, respectively. Variable activity was observed with metal ions and among inhibitors, chaotropic agents and organic solvents; phenyl glyoxal, potassium iodide, and butanol inhibited enzyme activity, respectively, while the enzyme activity was not majorly influenced by EDTA, urea, ethanol, and hexane. APF1 phytase treatment was found effective in dephytinization of flour biofortified wheat genotypes. Maximum decrease in phytic acid content was noticed in genotype MB-16-1-4 (89.98%) followed by PRH3-30-3 (82.32%) and PRH3-43-1 (81.47%). Overall, the study revealed that phytase from Aspergillus aculeatus APF1 could be effectively used in food and feed processing industry for enhancing nutritional value of food.

Phytases from Enterobacter and Serratia species with desirable characteristics for food and feed applications.

Phytases are enzymes of great industrial importance with wide range of applications in animal and human nutrition. These catalyze the hydrolysis of phosphomonoester bonds in phytate, thereby releasing lower forms of myo-inositol phosphates and inorganic phosphate. Addition of phytase to plant-based foods can improve its nutritional value and increase mineral bioavailability by decreasing nutritional effect of phytate. In the present investigation, 43 phytase positive bacteria on PSM plates were isolated from different sources and characterized for phytase activity. On the basis of phytase activity and zone of hydrolysis, two bacterial isolates (PSB-15 and PSB-45) were selected for further characterization studies, i.e., pH and temperature optima and stability, kinetic properties and effect of modulators. The phytases from both isolates were optimally active at the pH value from 3 to 8 and in the temperature range of 50-70 °C. Further, the stability of isolates was good in the pH range of 3.0-8.0. Much variation was observed in temperature and storage stability, responses of phytases to metal ions and modulators. The K m and V max values for PSB-15 phytase were 0.48 mM and 0.157 µM/min, while for PSB-45 these were 1.25 mM and 0.140 µM/min, respectively. Based on 16S rDNA gene sequence, the isolates were identified as Serratia sp. PSB-15 (GenBank Accession No. KR133277) and Enterobacter cloacae strain PSB-45 (GenBank Accession No. KR133282). The novel phytases from these isolates have multiple characteristics of high thermostability and good phytase activity at desirable range of pH and temperature for their efficient use in food and feed to facilitate hydrolysis of phytate-metal ion complex and in turn, increased bioavailability of important metal ions to monogastric animals.

Transfer of useful variability of high grain iron and zinc from Aegilops kotschyi into wheat through seed irradiation approach.

PURPOSE: To transfer the 2S chromosomal fragment(s) of Aegilops kotschyi (2S(k)) into the bread wheat genome which could lead to the biofortification of wheat with high grain iron and zinc content. MATERIALS AND METHODS: Wheat-Ae. kotschyi 2A/2S(k) substitution lines with high grain iron and zinc content were used to transfer the gene/loci for high grain Fe and Zn content into wheat using seed irradiation approach. RESULTS: Bread wheat plants derived from 40 krad-irradiated seeds showed the presence of univalents and multivalents during meiotic metaphase-I. Genomic in situ hybridization analysis of seed irradiation hybrid F2 seedlings showed several terminal and interstitial signals indicated the introgression of Ae. kotschyi chromosome segments. This proves the efficacy of seed radiation hybrid approach in gene transfer experiments. All the radiation-treated hybrid plants with high grain Fe and Zn content were analyzed with wheat group 2 chromosome-specific polymorphic simple sequence repeat markers to identify the introgression of small alien chromosome fragment(s). CONCLUSION: Radiation-induced hybrids showed more than 65% increase in grain iron and 54% increase in Zn contents with better harvest index than the elite wheat cultivar WL711 indicating effective and compensating translocations of 2S(k) fragments into wheat genome.

Bioavailability of iron from wheat aegilops derivatives selected for high grain iron and protein contents.

A coupled in vitro digestion/Caco-2 model was employed to assess iron bioavailability from wheat Aegilops derivatives selected for high iron and protein contents. The iron content in wheat genotypes used in this study correlated to a great extent with both protein (r = 0.80) and phytate (r = 0.68) contents. The iron bioavailability was based on Caco-2 cell ferritin formation from cooked digests of these derivatives (relative to WL711 control) and correlated positively with dialyzable iron (r = 0.63) and total iron content (r = 0.38) but not with the phytate content. The apparently decreased phytate/iron molar ratios, however, correlated negatively (r = -0.42) with the iron bioavailability, justifying the utilization of these parameters in biofortification programs. Iron bioavailability in the derivatives increased up to 1.5-fold, corresponding to a 1.5-2.2-fold increase observed in iron content over control. These data suggest that biofortification for iron proportionately leading to higher iron bioavailability will be the most feasible and cost-effective approach to combat micronutrient deficiency.

Negative effect of chromosome 1A on dough strength shown by modification of 1D addition in durum wheat (Triticum durum).

A monosomic addition line of Aegilops tauschii chromosome 1D in Triticum durum cv. PBW114 was produced in 1990. This line was self-pollinated and maintained for several generations while following the presence of chromosome 1D carrying the gene for red glume color. Cytological analysis indicated that two of the three derivative lines had substitution of chromosome 1D for 1A and another had substitution of chromosome 1D for 1B. One of these lines carried a pair of small chromosomes in addition to the 1D chromosome. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the derived lines showed the presence of high-molecular-weight (HMW) glutenin encoded by the Glu-D1 locus. The small chromosome found in one of the lines had nearly regular pairing and transmission to daughter nuclei. Fluorescent in situ hybridization (FISH) and analysis of molecular markers indicated that the small chromosome was derived from the short arm of chromosome 1A and carried the Glu-A3 locus. Microsatellite mapping based on the deletion bin map revealed that the small chromosome had terminal deletions on both the terminal and centromeric sides. The line with the small chromosome showed improvement of the sodium dodecyl sulfate (SDS)-sedimentation value as compared to parent durum. However, the increase in SDS-sedimentation value was more significant in the substitution line of chromosome 1D for 1A without the small chromosome. These facts suggest a negative effect of the Glu-A3 locus on dough strength. The sequence of the Glu-D1 locus from these lines showed that the HMW glutenin subunits were Ae. tauschii specific 2(t) + T2, which were previously found to be associated with poor rheological properties and bread loaf volume in synthetic hexaploid wheat by other workers. Thus, the significant improvement in the SDS-sedimentation value of the substitution line of 1D for 1A suggests that the absence of the negative effect of chromosome 1A on quality is more important than the presence of Glu-D1 of Ae. tauschii.