Physiological and genomic evidence supports the role of Serratia quinivorans PKL:12 as a biopriming agent for the biohardening of micropropagated Picrorhiza kurroa plantlets in cold regions.

The medicinal herb, Picrorhiza kurroa Royle ex Benth has become endangered because of indiscriminate over-harvesting. Although micropropagation has been attempted for mass propagation of the plant, survival of in vitro plantlets under green house/open field poses a major challenge. Biopriming of micropropagated plantlets with plant growth-promoting rhizobacteria (PGPR) are among the successful methods to combat this problem. Serratia quinivorans PKL:12 was the best-characterized PGPR from rhizospheric soil of P. kurroa as it increased the vegetative growth and survival of the micropropagated plantlets most effectively. Complete genome (5.29 Mb) predicted genes encoding proteins for cold adaptation and plant growth-promoting traits in PKL:12. Antibiotic and biosynthetic gene cluster prediction supported PKL:12 as a potential biocontrol agent. Comparative genomics revealed 226 unique genes with few genes associated with plant growth-promoting potential. Physiological and genomic evidence supports S. quinivorans PKL:12 as a potential agent for bio-hardening of micropropagated P. kurroa plantlets in cold regions.

De novo transcriptome provides insights into the growth behaviour and resveratrol and trans-stilbenes biosynthesis in Dactylorhiza hatagirea - An endangered alpine terrestrial orchid of western Himalaya.

This is the first report on de novo transcriptome of Dactylorhiza hatagirea, a critically-endangered, terrestrial orchid of alpine Himalayas. The plant is acclaimed for medicinal properties but little is known about its secondary-metabolites profile or cues regulating their biosynthesis. De novo transcriptome analysis was therefore, undertaken to gain basic understanding on these aspects, while circumventing the acute limitation of plant material availability. 65,384 transcripts and finally, 37,371 unigenes were assembled de novo from a total of 236 million reads obtained from shoot, tuber and leaves of the plant. Dominance of differentially-expressing-genes (DEGs) related to cold-stress-response and plant-hormone-signal-transduction; and those involved in photosynthesis, sugar-metabolism and secondary-metabolite-synthesis provided insights into carbohydrate-partitioning in the plant during its preparation for freezing winter at natural habitat. DEGs of glucomannan, ascorbic acid, carotenoids, phylloquinone/naphthoquinones, indole alkaloids, resveratrol and stilbene biosynthesis revealed the secondary-metabolite profile of D. hatagirea. UHPLC results confirmed appreciable amounts of resveratrol and trans-stilbene in D. hatagirea tubers, for the first time. Expression analysis of 15 selected genes including those of phenylpropanoid pathway confirmed the validity of RNA-seq data. Opportunistic growth, temperature- and tissue-specific-differential-expression of secondary metabolite biosynthesis and stress tolerant genes were confirmed using clonal plants growing at 8, 15 and 25 °C.

Proteome Analysis of the Gametophytes of a Western Himalayan Fern Diplazium maximum Reveals Their Adaptive Responses to Changes in Their Micro-Environment.

Ferns have survived changing habitats and environmental extremes of different eras, wherein, the exploratory haploid gametophytes are believed to have played a major role. Therefore, the proteome of in vitro grown gametophytes of a temperate Himalayan fern, Diplazium maximum in response to 0 (G0), 1 (G1), and 3% (G3) sucrose was studied. A total of 110 differentially abundant protein spots (DAPs) were obtained. Among these, only 67 could be functionally categorized as unique proteins involved in various metabolic processes. Calcium dependent proteins, receptor like kinases, G proteins, proteins related to hormonal signaling and their interaction with other pathways, and regulatory proteins were recorded indicating the involvement of five different signaling pathways. DAPs involved in the activation of genes and transcription factors of signaling and transduction pathways, transport and ion channels, cell-wall and structural proteins, defense, chaperons, energy metabolism, protein synthesis, modification, and turnover were identified. The gametophytes responded to changes in their micro-environment. There was also significant increase in prothallus biomass and conversion of two-dimensional prothalli into three-dimensional prothallus clumps at 3% sucrose. The three-D clumps had higher photosynthetic surface area and also closer proximity for sexual reproduction and sporophyte formation. Highest accumulation of proline, enhanced scavenging of reactive oxygen species (ROS) and DAPs of mostly, abiotic stress tolerance, secondary metabolite synthesis, and detoxification at 3% sucrose indicated an adaptive response of gametophytes. Protein Protein Interaction network and Principal Component analyses, and qRT-PCR validation of genes encoding 12 proteins of various metabolic processes indicated differential adjustment of gametophytes to different levels of sucrose in the culture medium. Therefore, a hypothetical mechanism was proposed to show that even slight changes in the micro-environment of D. maximum gametophytes triggered multiple mechanisms of adaptation. Many DAPs identified in the study have potential use in crop improvement and metabolic engineering programs, phytoremediation and environmental protection.

In vitro flowering associated protein changes in Dendrocalamus hamiltonii.

In Dendrocalamus hamiltonii, conversion of vegetative meristem to a floral meristem was successfully achieved on flower induction medium. A total of 128 differentially expressed proteins were evidenced by 2DE in floral meristem protein profiles. Analysis of 103 proteins through PMF revealed change in abundance in the content of 79 proteins, disappearance and new appearance in the content of 7 and 17 proteins, respectively. MS/MS and subsequent homology search identified 65 proteins that were involved in metabolism (22 proteins), regulatory (11 proteins), signaling and transportation (12 proteins), stress (6 proteins), flowering (8 proteins), and unknown functions (6 proteins). The data suggested that change in metabolism related proteins might be providing nutrient resources for floral initiation in D. hamiltonii. Further, interactive effects of various proteins like bHLH145, B-4c transcription factors (heat stress transcription factor), maturase K, MADS box, zinc finger proteins, and scarecrow-like protein 21 (flowering related), a key enzyme of ethylene biosynthesis SAMS (S-adenosylmethionine synthase) and aminocyclopropane-1-carboxylate synthase, improved calcium signaling related proteins (CML36), and change in phytohormone related proteins such as phosphatase proteins (2c3 and 2c55), which are the positive regulators of gibberellic acid and phytochrome regulation related proteins (DASH, LWD1) might be the possible major regulators of floral transition in this bamboo.

In vitro flowering--a system for tracking floral organ development in Dendrocalamus hamiltonii Nees et Arn. ex Munro.

Dendrocalamus hamiltonii plants are slender and tall (15-25 m) thereby, rendering tagging, sampling and tracking the development of flowers difficult. Therefore, a reproducible system of in vitro flowering was established for tracking the stages of flower development. MS medium supplemented with 2.22 microM 6-benzylaminopurine, 1.23 microM indole-3-butyric acid and 2% sucrose was optimized as the flower induction medium (FIM) wherein 28 and 42 days were required for the development of gynoecium and androecium, respectively. Six distinct stages of in vitro flower development were identified, and the flowers were comparable with that of in planta sporadic flowers. Pollen viability of the in vitro flowers was higher than those of in planta ones. The in vitro system developed in the present study facilitates easy tracking of different stages of flower development under controlled environmental conditions. It can also be used for medium- or long-term storage of pollens and manipulation of in vitro fertilization.

Mitral annulus displacement measured by two-dimensional speckle tracking imaging to assess the left ventricular longitudinal systolic function in coronary heart disease.

AIM: To assess the clinical utility of measuring mitral annulus displacement (MAD) by two-dimensional speckle tracking for the rapid evaluation of left ventricular longitudinal systolic function in patients with coronary heart disease (CHD). METHODS: Left ventricular longitudinal systolic function was evaluated by MAD using speckle tracking echocardiography in 30 healthy volunteers (controls) as well as in 30 patients with mild, 30 patients with moderate, and 30 patients with severe CHD. All participants had their apical four-chamber and two-chamber view echocardiographic images recorded. Left ventricular end-diastolic volume, left ventricular end-systolic volume, and left ventricular ejection fraction were calculated by the biplane Simpson method. MAD of interventricular septum (MADsep), left ventricular lateral wall (MADlat), and the middle point of the mitral annulus (MADmid) were measured offline with speckle-tracking echocardiography. MADmid% was defined as MADmid divided by left ventricular end-diastolic diameter. RESULTS: MADmid% was 16.9 ± 1.9, 11.8 ± 3.2, 11.8 ± 2.9, and 10.3 ± 3.6, respectively, in controls and in patients with mild, moderate, and severe CHD. All MAD indexes were lower in patients with moderate or severe CHD than in controls or patients with mild CHD. CONCLUSIONS: MAD is an early and rapid index for the assessment of left ventricular longitudinal systolic function in patients with different degrees of coronary artery disease.