The Plant-Rhizobial Symbiotic Interactions Provide Benefits to the Host beyond Nitrogen Fixation That Promote Plant Growth and Productivity

Rhizobial symbiotic interactions are known for nitrogen fixation, providing commercial crops and other plants with self-sufficiency in nitrogen requirements. An enormous contribution from nitrogen fixation is vital to the global nitrogen cycle. The symbiotic nitrogen reduces the carbon footprint of crop cultivation, which underlines its importance in agricultural sustainability. Extensive research efforts have been made to understand the symbiotic relationship at molecular, physiological, and ecological levels. This led to the isolation and modification of symbiotic strains for enhanced nitrogen efficiency. During the evaluation of strains for nitrogen fixation in exchange for supporting the bacterium in terms of space and resources, it has been observed that the accrued benefits to the host plants extend well beyond the nitrogen fixation. The symbiotic interaction has been advantageous to the host for better growth and development, tolerating a stressful environment, and even keeping the pathogenic microbial enemies at bay. Additionally, it enabled the availability of the mineral nutrients, which otherwise were inaccessible to the host. In this chapter, we bring together the information with a focus on the role of rhizobial symbiotic interactions that promote plant growth and productivity through phytohormone synthesis, by facilitating the availability of mineral nutrients, and by improving the plant tolerance to sub-optimal growth conditions

Physical mapping of wheat and rye expressed sequence tag-simple sequence repeats on wheat chromosomes

Six hundred and seventy two loci belonging to 275 expressed sequence tag-simple sequence repeats [EST-SSRs, including 93 wheat (Triticum aestivum L.) and 182 rye (Secale cereale L.) EST-SSRs] were physically mapped on 21 wheat chromosomes. The mapping involved two approaches, the wet-lab approach involving use of deletion stocks and the in silico approach involving matching with ESTs that were previously mapped. The number of loci per EST-SSR mapped using the in silico approach was almost double the number of loci mapped using the wet-lab approach (using deletion stocks). The distribution of loci on the three subgenomes, on the seven homoeologous groups and on the 21 individual chromosomes was nonrandom (P « 0.01). Long arms had disproportionately (relative to the difference in DNA content) higher number of loci, with more loci mapped to the distal regions of chromosome arms. A fairly high proportion of EST-SSRs had multiple loci, which were largely (81%) homoeoloci. Rye EST-SSRs showed a high level of transferability (≈77%) to the wheat genome. Putative functions were assigned to 216 SSR-containing ESTs through homology searches against the protein database. As many as 104 SSR-containing ESTs (a subset of the above ESTs) were also mapped to the 12 rice chromosomes, which corresponded with the known homology between wheat and rice chromosomes. These physical maps of EST-SSRs should prove useful for comparative genomics, gene tagging, fine mapping, and cloning of genes and QTLs. Dna-based molecular markers, particularly SSRs, have been developed and mapped on chromosomes in a variety of crop plants. In bread wheat, genetic and physical mapping of SSRs has been an ongoing exercise, and, to date, ≈2450 SSRs (1 SSR 1.63 cM-1) have been genetically mapped (for details see Torada et al., 2006) and ≈1320 SSRs (62 SSRs chromosome-1) have been physically mapped (for details see Goyal et al., 2005). With a genome size of ≈16 000 Mbp, it is evident that despite concerted efforts, the density of mapped SSRs in bread wheat remains relatively low and continued efforts are needed to increase the density of these SSRs on available genetic and physical maps. In recent years, emphasis has also shifted from genomic SSRs to EST-SSRs due to the availability of very large databases of ESTs from all of the cereals including bread wheat. Consequently, the number of EST-SSRs in cereals now includes 43 598 from bread wheat (Peng and Lapitan, 2005), 16 917 from rice and 184 from rye (La Rota et al., 2005; Hackauf and Wehling, 2002). The genetic mapping of these EST-SSRs is difficult due to a low level of polymorphism, as a result of their conserved nature. Physical mapping of these EST-SSRs in wheat is equally difficult due to the occurrence of homoeoloci exhibiting no polymorphism. This has discouraged wheat researchers from undertaking a large-scale project to genetically or physically map wheat EST-SSRs although genetic mapping of 325 EST-SSRs (Gao et al., 2004; Nicot et al., 2004; Yu et al., 2004) and physical mapping of 305 EST-SSRs was recently undertaken (Yu et al., 2004; Zhang et al., 2005; Peng and Lapitan, 2005). We previously reported genetic mapping of 58 and physical mapping of 270 genomic SSRs (Gupta et al., 2002; Goyal et al., 2005). The present study is an extension of our earlier studies on physical mapping of SSRs and involved both wet-lab and in silico approaches, leading to the successful mapping of as many as 672 loci. The in silico approach allowed mapping of twice the number of loci (per EST-SSR) mapped using wet-lab analysis

A field-grown transgenic tomato line expressing higher levels of polyamines reveals legume cover crop mulch-specific perturbations in fruit phenotype at the levels of metabolite profiles, gene expression, and agronomic characteristics

Genetic modification of crop plants to introduce desirable traits such as nutritional enhancement, disease and pest resistance, and enhanced crop productivity is increasingly seen as a promising technology for sustainable agriculture and boosting food production in the world. Independently, cultural practices that utilize alternative agriculture strategies including organic cultivation subscribe to sustainable agriculture by limiting chemical usage and reduced tillage. How the two together affect fruit metabolism or plant growth in the field or whether they are compatible has not yet been tested. Fruit-specific yeast S-adenosylmethionine decarboxylase (ySAMdc) line 579HO, and a control line 556AZ were grown in leguminous hairy vetch (Vicia villosa Roth) (HV) mulch and conventional black polyethylene (BP) mulch, and their fruit analysed. Significant genotype×mulch-dependent interactions on fruit phenotype were exemplified by differential profiles of 20 fruit metabolites such as amino acids, sugars, and organic acids. Expression patterns of the ySAMdc transgene, and tomato SAMdc, E8, PEPC, and ICDHc genes were compared between the two lines as a function of growth on either BP or HV mulch. HV mulch significantly stimulated the accumulation of asparagine, glutamate, glutamine, choline, and citrate concomitant with a decrease in glucose in the 556AZ fruits during ripening as compared to BP. It enables a metabolic system in tomato somewhat akin to the one in higher polyamine-accumulating transgenic fruit that have higher phytonutrient content. Finally, synergism was found between HV mulch and transgenic tomato in up-regulating N:C indicator genes PEPC and ICDHc in the fruit

Development of a facile antibody–drug conjugate platform for increased stability and homogeneity† †Electronic supplementary information (ESI) available: Synthetic schemes and characterization data, experimental procedures, Fig. S1 and S2. See DOI: 10.1039/c6sc05149a Click here for additional data file.

Despite the advances in the design of antibody–drug conjugates (ADCs), the search is still ongoing for novel approaches that lead to increased stability and homogeneity of the ADCs. We report, for the first time, an ADC platform technology using a platinum(ii)-based linker that can re-bridge the inter-chain cysteines in the antibody, post-reduction. The strong platinum–sulfur interaction improves the stability of the ADC when compared with a standard maleimide-linked ADC thereby reducing the linker–drug exchange with albumin significantly. Moreover, due to the precise conserved locations of cysteines, both homogeneity and site-specificity are simultaneously achieved. Additionally, we demonstrate that our ADCs exhibit increased anticancer efficacy in vitro and in vivo. The Pt-based ADCs can emerge as a simple and exciting proposition to address the limitations of the current ADC linker technologies

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Molecular Biology in the Improvement of Biological Nitrogen Fixation by Rhizobia and Extending the Scope to Cereals

The contribution of biological nitrogen fixation to the total N requirement of food and feed crops diminished in importance with the advent of synthetic N fertilizers, which fueled the “green revolution”. Despite being environmentally unfriendly, the synthetic versions gained prominence primarily due to their low cost, and the fact that most important staple crops never evolved symbiotic associations with bacteria. In the recent past, advances in our knowledge of symbiosis and nitrogen fixation and the development and application of recombinant DNA technology have created opportunities that could help increase the share of symbiotically-driven nitrogen in global consumption. With the availability of molecular biology tools, rapid improvements in symbiotic characteristics of rhizobial strains became possible. Further, the technology allowed probing the possibility of establishing a symbiotic dialogue between rhizobia and cereals. Because the evolutionary process did not forge a symbiotic relationship with the latter, the potential of molecular manipulations has been tested to incorporate a functional mechanism of nitrogen reduction independent of microbes. In this review, we discuss various strategies applied to improve rhizobial strains for higher nitrogen fixation efficiency, more competitiveness and enhanced fitness under unfavorable environments. The challenges and progress made towards nitrogen self-sufficiency of cereals are also reviewed. An approach to integrate the genetically modified elite rhizobia strains in crop production systems is highlighted

Characterization of natural antimony resistance in Leishmania Donovani isolates

Clinical resistance to pentavalent antimonial compounds has long been recognized as a major problem in the treatment of visceral leishmaniasis in India. However, mechanisms of natural resistance are unclear. In this study, we observed that Leishmania donovani clinical isolates not responsive to sodium stibogluconate showed resistance to antimony treatment in both in vitro and in vivo laboratory conditions. The resistant isolates have increased levels of intracellular thiols. This increase in thiol levels was not mediated by the amplification of ?-glutamylcysteine synthetase, but was accompanied by amplification of trypanothione reductase and an intracellular ATP-binding cassette transporter gene MRPA. The resistance of parasites to antimony could be reversed by the glutathione biosynthesis-specific inhibitor, buthionine sulfoximine, which resulted in increased drug susceptibility. These results suggest the possible role of thiols and MRPA in antimony resistance in field isolates

Enhanced flux of substrates into polyamine biosynthesis but not ethylene in tomato fruit engineered with yeast S-adenosylmethionine decarboxylase gene

S-adenosylmethionine (SAM), a major substrate in 1-C metabolism is a common precursor in the biosynthetic pathways of polyamines and ethylene, two important plant growth regulators, which exhibit opposing developmental effects, especially during fruit ripening. However, the flux of various substrates including SAM into the two competing pathways in plants has not yet been characterized. We used radiolabeled 14C-Arg, 14C-Orn, L-[U-14C]Met, 14C-SAM and 14C-Put to quantify flux through these pathways in tomato fruit and evaluate the effects of perturbing these pathways via transgenic expression of a yeast SAM decarboxylase (ySAMDC) gene using the fruit ripening-specific promoter E8. We show that polyamines in tomato fruit are synthesized both from Arg and Orn; however, the relative contribution of Orn pathway declines in the later stages of ripening. Expression of ySAMDC reversed the ripening associated decline in spermidine (Spd) and spermine (Spm) levels observed in the azygous control fruit. About 2- to 3-fold higher levels of labeled-Spd in transgenic fruit (556HO and 579HO lines) expressing ySAMDC confirmed the enzymatic function of the introduced gene. The incorporation of L-[U-14C]Met into Spd, Spm, ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) was used to determine Met-flux into these metabolites. The incorporation of 14C-Met into Spd/Spm declined during ripening of the control azygous fruit but this was reversed in fruits expressing ySAMDC. However, incorporation of 14C-Met into ethylene or ACC during ripening was not altered by the expression of ySAMDC in the fruit. Taken together these results show that: (1) There is an inverse relationship between the production of higher polyamines and ethylene during fruit ripening, (2) the inverse relationship between higher polyamines and ethylene is modulated by ySAMDC expression in that the decline in Spd/Spm during fruit ripening can be reversed without significantly altering ethylene biosynthesis, and (3) cellular flux of SAM in plants is homeostatically regulated based on its demand for competing pathways

<i>msrA3</i>-transgenic plants display delay in dark-induced senescence.

<p>(a) Detached compound leaves (5^th to 6^th from top) of five week-old plants were incubated in the dark on moist filter paper in Petri dishes at room temperature and were photographed after 5 days incubation in dark. The Petri dishes were wrapped in aluminum foil to create dark conditions. The experiment was repeated at least three times and the representative results are shown here. (b) Percent retention of chlorophyll in wild-type and transgenic plants. The leaves were sampled for analysis at indicated times after incubation in darkness. Bars represent means <u>+</u> SE (n=3). (c) Expression profile of genes involved in HR and associated senescence in wild-type and <i>msrA3</i>-transgenic plant leaves kept in the dark (as stated in section [a]) at room temperature for the indicated times. Each lane was loaded with 20 μg of total RNA. Ethidium bromide stained rRNA served as loading control.</p

The development of floral buds is delayed in the <i>msr</i>A3-transgenics (T3 and T26).

<p>(a) Wild-type (WT) plant with the earliest emergence of a floral bud at 20 days after germination. The circles mark the terminal shoot of transgenics showing no signs of flower bud initiation. The buds were observed only on the main shoots from three replicated pots each having 3 plants grown in a growth chamber at day/night temperatures of 28⁰/22⁰C and 16/8 h light/dark cycle with light intensity of 300 µM quanta.m^-2.s^-1; (b) Number of newly appeared buds after indicated days of germination. The buds were counted from six replicated pots each having 4-5 plants totaling 28. The plants were grown under natural light in green house with 16 h day length, and night and day temperature varied between12-33°C.</p