Stable Plants Research Articles

Integral Control of Stable MIMO Nonlinear Systems with Input Constraints

Abstract This paper presents a novel anti-windup integral controller for a stable multi-input multi-output nonlinear plant. We use tools from projected dynamical systems theory to force the controller state (and thus its output) to remain in a “permitted” (compact and convex) region, regardless of the plant behaviour. Then, using results from singular perturbation theory, we find a sufficient condition on the controller gain ensuring, under suitable monotonicity assumptions on the plant, closed-loop stability and reference tracking for a feasible set of constant references.

TDZ induced plant regeneration from immature cotyledons of Pterocarpus marsupium Roxb. and validation of genetic homogeneity using ISSR markers

Pterocarpus marsupium Roxb. (Indian kino) is a threatened leguminous tree species well known for its therapeutic properties. A genetically stable plant regeneration system in P.marsupium has been developed using immature cotyledon (IC) explants excised from 12-days old in vitro grown seedlings. ICs were cultured on Murashige and Skoog (MS) media augmented with various concentrations of plant growth regulators to induce multiple shoots. Among the various concentrations and combinations used, MS medium augmented with thidiazuron (TDZ; 4.45 µM) and naphthalene acetic acid (NAA; 1.07 µM) showed the highest regeneration frequency (95.83%) with a maximum number of adventitious shoot buds (25.4 ± 0.55) with 22.7 shoots per explant. The harmful effects of extended exposure to TDZ at long-term regimen was prevented by transferring the organogenic cultures to MS medium fortified with 6- benzylaminopurine (BAP) for further multiplication and elongation. The highest number of shoots (36.0 ± 0.74) and shoot length (5.8 ± 0.10) were observed on 4.44 µM BAP augmented media after the third subculture. The elongated shoots were treated for 24 h with 19.60 µM indole-3-butyric acid (IBA) solution and rooted on half strength MS medium amended with 2.85 µM IBA. The plantlets were acclimatized successfully in the greenhouse with a 90% survival rate. Genetic homogeneity of regenerants were analyzed by inter simple sequence repeat (ISSR) markers. The comparative amplified products across the in vitro developed plantlets and the mother plant showed good genetic consistency. The protocol mentioned herein would be beneficial for raising genetically uniform plantlets and commercial cultivation of P. marsupium.

Agrobacterium T-DNA integration in somatic cells does not require the activity of DNA polymerase θ.

Integration of Agrobacterium tumefaciens transferred DNA (T-DNA) into the plant genome is the last step required for stable plant genetic transformation. The mechanism of T-DNA integration remains controversial, although scientists have proposed the participation of various nonhomologous end-joining (NHEJ) pathways. Recent evidence suggests that in Arabidopsis, DNA polymerase θ (PolQ) may be a crucial enzyme involved in T-DNA integration. We conducted quantitative transformation assays of wild-type and polQ mutant Arabidopsis and rice, analyzed T-DNA/plant DNA junction sequences, and (for Arabidopsis) measured the amount of integrated T-DNA in mutant and wild-type tissue. Unexpectedly, we were able to generate stable transformants of all tested lines, although the transformation frequency of polQ mutants wasc. 20% that of wild-type plants. T-DNA/plant DNA junctions from these transformed rice and Arabidopsis polQ mutants closely resembled those from wild-type plants, indicating that loss of PolQ activity does not alter the characteristics of T-DNA integration events. polQ mutant plants show growth and developmental defects, perhaps explaining previous unsuccessful attempts at their stable transformation. We suggest that either multiple redundant pathways function in T-DNA integration, and/or that integration requires some yet unknown pathway.

Sine Cosine Algorithm Assisted FOPID Controller Design for Interval Systems Using Reduced-Order Modeling Ensuring Stability

The focus of present research endeavor was to design a robust fractional-order proportional-integral-derivative (FOPID) controller with specified phase margin (PM) and gain cross over frequency (ωgc) through the reduced-order model for continuous interval systems. Currently, this investigation is two-fold: In the first part, a modified Routh approximation technique along with the matching Markov parameters (MPs) and time moments (TMs) are utilized to derive a stable reduced-order continuous interval plant (ROCIP) for a stable high-order continuous interval plant (HOCIP). Whereas in the second part, the FOPID controller is designed for ROCIP by considering PM and ωgc as the performance criteria. The FOPID controller parameters are tuned based on the frequency domain specifications using an advanced sine-cosine algorithm (SCA). SCA algorithm is used due to being simple in implementation and effective in performance. The proposed SCA-based FOPID controller is found to be robust and efficient. Thus, the designed FOPID controller is applied to HOCIP. The proposed controller design technique is elaborated by considering a single-input-single-output (SISO) test case. Validity and efficacy of the proposed technique is established based on the simulation results obtained. In addition, the designed FOPID controller retains the desired PM and ωgc when implemented on HOCIP. Further, the results proved the eminence of the proposed technique by showing that the designed controller is working effectively for ROCIP and HOCIP.

Protein phosphatase NtPP2C2b and MAP kinase NtMPK4 act in concert to modulate nicotine biosynthesis.

Protein phosphatases (PPs) and protein kinases (PKs) regulate numerous developmental, defense, and phytohormone signaling processes in plants. However, the underlying regulatory mechanism governing biosynthesis of specialized metabolites, such as alkaloids, by the combined effects of PPs and PKs, is insufficiently understood. Here, we report the characterization of a group B protein phosphatase type 2C, NtPP2C2b, that likely acts upstream of the NICOTINE2 locus APETALA 2/Ethylene Response Factors (AP2/ERFs), to regulate nicotine biosynthesis in tobacco. Similar to the nicotine pathway genes, NtPP2C2b is highly expressed in roots and induced by jasmonic acid (JA). Overexpression of NtPP2C2b in transgenic hairy roots or stable transgenic tobacco plants repressed nicotine pathway gene expression and reduced nicotine accumulation. Additionally, transient overexpression of NtPP2C2b, together with the NtERF221, repressed transactivation of the quinolinate phosphoribosyltransferase promoter in tobacco cells. We further demonstrate that the JA-responsive tobacco mitogen-activated protein kinase (MAPK) 4 interacts with NtPP2C2b in yeast and plant cells. Conditional overexpression of NtMPK4 in tobacco hairy roots up-regulated nicotine pathway gene expression and increased nicotine accumulation. Our findings suggest that a previously uncharacterized PP-PK module acts to modulate alkaloid biosynthesis, highlighting the importance of post-translational control in the biosynthesis of specialized plant metabolites.

Pleistocene freshwater environments of Poland: a comprehensive study of fish assemblages based on a multi‐proxy approach

The results of a comprehensive study of Pleistocene fish‐bearing localities from the territory of Poland are presented. Fish remains came from lacustrine deposits and cave sites representing a time‐span from the late Early Pleistocene until the Early Holocene. Here we present the taxonomic composition and species diversity of all known fish assemblages from Poland. In total, 23 species belonging to 20 genera of eight families were identified. Cyprinids were the most taxonomically diverse, followed by salmonids and percids. Other families were each represented only by a single species. The roach, rudd, bream, pike and perch were the most common components of the Pleistocene fish assemblages of Poland. These assemblages are clearly divided into two groups representing palaeolakes and cave sites. A significant taxonomic similarity is observed between fish assemblages from Poland and those of Chibanian and Late Pleistocene ages from central and eastern Europe. Pleistocene environments and climates of Poland are characterized using a multi‐proxy approach (analysis of stable isotopes, plant macro‐remains, pollen, molluscs and cladocerans). Obtained data allowed us to estimate a number of parameters of Pleistocene water bodies of the Polish Lowland (reservoir type, trophism, oxygen content, depth and water level changes, bottom character, and flow speed). The formation of freshwater fish assemblages was influenced by dynamic changes in the hydrographic network, and climatic changes during the Pleistocene and Holocene. Their development was favoured by the formation of lakes after melting of the ice sheet.

Analysis and Synthesis of Low-Gain Integral Controllers for Nonlinear Systems

Relaxed conditions are given for stability of a feedback system consisting of an exponentially stable multi-input multi-output nonlinear plant and an integral controller. Roughly speaking, it is shown that if the composition of the plant equilibrium input-output map and the integral feedback gain is infinitesimally contracting, then the closed-loop system is exponentially stable if the integral gain is sufficiently low. The main result is illustrated with an application arising in frequency control of ac power systems. We demonstrate how the contraction condition can be checked computationally via semidefinite programming, and how integral gain matrices can be synthesized via convex optimization to achieve robust \mathscrL <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> performance in the presence of nonlinearity and uncertainty.

The breeding method and its influence on the intensification of the tomato selection process

Relevance. The article presents the results of using three different selection methods to obtain new forms of tomato that combine high productivity with resistance to high-temperature stress factor.Methods. Eleven hybrid combinations were used as experimental material. Selections were made starting with the offspring of the F1 generation. Three different methodological approaches were used: 1 – step-by-step alternation of sporophytic-gametophytic selections under high temperature pressure; 2 – only gametophytic selection using heat-treated pollen for pollination; 3 – traditional selections based on a high value of indicators of the studied traits. In each of the variants of the experiment, we studied: the heat resistance of offspring at different stages of ontogenesis – "sporophyte-gametophyte"; the duration of the vegetation period of plants; the fruit formation; the total yield. The offspring of the most stable and productive plants isolated from different hybrid combinations were combined in the populations in each variant of the experiment and a comparative analysis and evaluation of the effectiveness of the methods used was carried out.Results. It is shown that the combination of step-by-step sporophyte-gametophyte selections under the action of high-temperature regimes (43°C and 45°C), at early stages (F1–F3) lead to more stable, productive offspring in F5-F7 both for sporophyte and gametophyte traits, compared to using only gametophyte selections and stronger differences relative to the control variant. The gradual alternation of selections at different stages of ontogenesis also leads to the production of offspring with a shorter period of passage of the phenophases "seedling-flowering" – "floweringmaturation" and a stable manifestation of signs that determine the quality of the fruits. Using only gametophytic selections and traditional methods of selection was less effective than alternating sporophyte-gametophyte ones.Conclusions. The differentiating ability of the selection methods used and their influence on obtaining more resistance and productive genotypes was established. The advantage of the method of step-by-step alternation of sporophyte-gametophyte selections, which led to the production of new lines (118/1, 124/1, 133-11/1, 133-12/1, 134/1, 110/1 and 7/1) with high resistance at different stages of ontogenesis (sporophyte-gametophyte), shorter vegetation period and high fruit-forming capacity in three different years of research (2015-2017).

Regeneration of Viburnum dentatum L. from Alginate-Encapsulated Shoot Explants after Short-Term Cold Storage and Assessment of Genetic Stability Using ISSR Analysis

The present study demonstrates an efficient protocol for alginate encapsulation, interim cold storing of artificial seeds and conversion to genetically stable plants of Viburnum dentatum L. “Lucidum Aiton”. Explants of shoot tips and first-node segments, excised from in vitro-derived viburnum microshoots, were encapsulated in 2.5% sodium alginate mixed with liquid MS nutrient medium and hardened in 50 mM of calcium chloride producing solid, soft and uniform beads. These artificial seeds achieved 28.9% germination under light, forming 4.3 microshoots per bead. However, with 100 mM of calcium chloride for hardening, the beads were firm and of a uniform globular shape and suitable for handling and exhibited a germination response of 48.9%. Encapsulated shoot tip explants of viburnum, which were stored at 4 °C for 4, 8 or 12 weeks, showed a gradual decline in regeneration response (73.3, 62.2, 51.1%, respectively), while non-encapsulated explants, stored under same conditions, did not survive after the fourth week of cold storage. Microshoots from cold-stored encapsulated explants, which were rooted in solid MS nutrient medium with 0.5 μΜ of Indole-3-acetic acid (IAA) and transplanted to a substrate of peat-perlite (3:1, v/v), acclimatized successfully after application of 75 or 50% shading, which was gradually reduced, and were established with minimum losses in a greenhouse. For the genetic stability of the artificial seed-derived plantlets and compared with the mother plant, an assessment was conducted using Inter Simple Sequence Repeats (ISSRs) analysis. The ISSR profiles proved the genetic uniformity and clonal stability of the regenerated plantlets and their genetic resemblance to the mother plant. The present regeneration procedure could be used as an alternative method for the micropropagation of V. dentatum.

Optimization of regeneration and Agrobacterium-mediated transformation of Stevia (Stevia rebaudiana Bertoni): a commercially important natural sweetener plant

Stevia rebaudiana Bertoni is a commercially important zero calorie natural-sweetener herb which produce sweet compounds known as steviol glycosides. Rising demands of steviol glycosides by food and beverage industries has led to an increase in its cultivation in various countries. Unfortunately, stevia cultivation faces 2–25% yield penalty due to weeds which further adds to its cultivation cost. To resolve this major challenge, Agrobacterium-mediated genetic transformation of in vitro derived stevia-nodal explants using herbicide resistance gene (bar) has been optimized, for the production of stable transgenic stevia plants. Several parameters including explant type, pre-incubation duration, acetosyringone (As) concentration, Agrobacterium cell density, Agro-inoculation duration, co-cultivation duration, selection regime and plant growth regulators (PGRs) combination and concentration, have been successfully optimized. Among the two types of explants used, nodal explants showed a higher regeneration response of 82.85%, with an average of 25 shoots/explant. The best PGRs combination and concentration for shoot-induction, shoot-elongation and root-induction was found to be 6-benzyladenine (1.0 mg l−1) + naphthalene acetic acid (0.5 mg l−1), gibberellic acid (1.0 mg l−1), and half-strength MS medium, respectively. The two-step selection (phosphinothricin) regime resulted in an average transformation efficiency of 40.48% with nodal explants. Molecular characterization of putative transformants through PCR, RT-PCR, qRT-PCR and Southern-blot hybridization confirmed the presence, stability, expression as well as copy number of bar gene respectively. Compared to the non-transgenic plants, the T0 transgenic plants successfully tolerated 8 mg l−1 glufosinate ammonium sprays. Thus, the optimized protocol can be useful for the introduction of other genes (inter-kingdom transfer) into stevia genome.

Use of Flow Cytometry for Producing Tetraploids in Red Clover

Abstract Sprouts of diploid red clover ‘Stendes Agrais’ were treated with 0.2% colchicine solution. The ploidy level of plants was detected by flow cytometry. Most of the treated plants were mixoploids and sterile. From progenies of fertile plants, mixoploids were selected that had a proportion of tetraploid cells higher than 60%. In the next generation, stable tetraploid plants were selected, and 49 lines were included in the breeding process.

A competence of embryo-derived tissues of tetraploid cultivated wheat species Triticum dicoccum and Triticum timopheevii for efficient and stable transgenesis mediated by particle inflow gun

BackgroundThe ability to engineer cereal crops by gene transfer technology is a powerful and informative tool for discovering and studying functions of genes controlling environmental adaptability and nutritional value. Tetraploid wheat species such as emmer wheat and Timopheevi wheat are the oldest cereal crops cultivated in various world areas long before the Christian era. Nowadays, these hulled wheat species are gaining new interest as donors for gene pools responsible for the improved grain yield and quality, tolerance for abiotic and biotic stress, resistance to pests and disease. The establishing of efficient gene transfer techniques for emmer and Timopheevi wheat may help in creation of modern polyploid wheat varieties.ResultsIn the present study, we describe a robust protocol for the production of fertile transgenic plants of cultivated emmer wheat (Russian cv. ‘Runo’) using a biolistic delivery of a plasmid encoding the gene of green fluorescent protein (GFP) and an herbicide resistance gene (BAR). Both the origin of target tissues (mature or immature embryos) and the type of morphogenic calli (white or translucent) influenced the efficiency of stable transgenic plant production in emmer wheat. The bombardment of nodular white compact calluses is a major factor allowed to achieve the highest transformation efficiency of emmer wheat (on average, 12.9%) confirmed by fluorescence, PCR, and Southern blot. In the absence of donor plants for isolation of immature embryos, mature embryo-derived calluses could be used as alternative tissues for recovering transgenic emmer plants with a frequency of 2.1%. The biolistic procedure based on the bombardment of immature embryo-derived calluses was also successful for the generation of transgenic Triticum timopheevii wheat plants (transformation efficiency of 0.5%). Most of the primary events transmitted the transgene expression to the sexual progeny.ConclusionThe procedures described here can be further used to study the functional biology and contribute to the agronomic improvement of wheat. We also recommend involving in such research the Russian emmer wheat cv. ‘Runo’, which demonstrates a high capacity for biolistic-mediated transformation, exceeding the previously reported values for different genotypes of polyploid wheat.

COURSE-KEEPING CONTROL FOR DIRECTIONALLY UNSTABLE LARGE TANKERS USING THE MIRROR-MAPPING TECHNIQUE

This study examines the course-keeping control of directionally unstable large oil tankers involving a pole in the right half plane. Treated as an unstable plant in control engineering, tankers are theoretically and experimentally investigated during the controller design process. First, the unstable plant is mirror-mapped to its corresponding stable minimum phase plant using the mirror-mapping technique, which enables an easy controller design. Then, a linear proportional-differential and a first-order filter controller is designed based on the closed-loop gain shaping algorithm, which requires only one controller parameter to be properly selected based on the system’s characteristics. Numerical simulation results confirmed that the designed controller can successfully stabilise an unstable plant subjected to external wind and wave disturbances. The controller designed with the proposed method is suitable for course-keeping control of directionally unstable large tankers. The controller design method is simple with an uncomplicated structure that can easily be implemented in engineering endeavours. Moreover, the rudder motion is small and soft.

Asymmetries in the Disturbance Compensation Methods for the Stable and Unstable First Order Plants

This paper analyzes the first-order and first-order time-delayed systems control approaches, focusing mainly on unstable systems. First, it discusses asymmetries between the disturbance observer-based (DOB) control with decoupled tracking and the disturbance rejection responses, stressing applications to stable and unstable plants. The paper analyzes some DOB-based control solutions for unstable systems which do not use internal closed-loop stabilization. The novelty of the paper is thorough study accompanied with a comprehensive explanation of the differences between two distinct approaches: the transfer-function- and the closed-loop-based feedforward control approach from the point of view of control constraints. It is clearly illustrated that the main cause of instability of DOB-based approaches, applied to unstable systems, is given by their effort to impose on the system the unstable dynamics of the chosen nominal process model. It is also shown that the closed-loop stability of the DOB-based control, applied to the unstable systems, can be restored by using the supervising reference model control (RMC). The main novelty of the proposed approach is that its eliminates the mentioned stability problems while maintaining the full functionality of the chosen control structures. RMC has so far only been implemented for generating a setpoint feedforward signal. However, by generalization of this approach for disturbance rejection, the methodology of DOB design, based on nominal models, can be extended to the control of unstable systems. Without the use of disturbance reference models, the interactions of the master stabilizer with disturbance compensation cannot be eliminated. Without the internal stabilization, the stable transients can only be achieved by designing controllers based on stable models, instead of unstable ones. The existing modifications of DOB-based schemes for unstable plants, proposed in some references, are shown to lead to traditional Proportional-Integrative (PI) control, thus losing all the advantages over the PI controllers. In all the considered structures, the role of integrating models is also emphasized.

PENGARUH ELISITOR Al3+ TERHADAP AKUMULASI DAIDZEIN DAN GENISTEIN PADA KALUS Phaseolus vulgaris L. Dan Phaseolus aureus Roxb.

The Estrogen was very beneficial hormone for men and women. Isoflavon compound was natural estrogen source which contain three main compound in chemical form, the aglycones, daidzein and genistein. They were found only 0,25% on plant, exclusively on leguminous plant. An alternative method have been required to overcome these problem, for example, through plant cell culture technic with heavy metal elicitor. The aims of this research were to study the effect of various concentration of Al3+ elicitor and age of culture for callus growth and accumulation of daidzein and genistein from in vitro Phaseolus vulgaris L. And Phaseolus aureus Roxb callus culture. Stable plant callus culture were established and subculture for three times approximately 4 weeks. The calli were elicitated by Al3+ elicitor with 0, 50, 150, 300 µM concentrations and incubated on 4, 6, and 8 weeks. Qualitative data were determined based in changed calli colour, TLC plate and value of Rf. Quantitative data were determined based in fresh and dry weight of the calli and the content of daidzein and genistein compound. Quantitative data were analized using Anova and continued with DMRT 95%. The results showed that the callus growth more influenced by age of culture. The sixth weeks led to higher fresh and dry weight. The Colour of Calli had been changed from white yellowness become brown at the eighth weeks. The texture of Calli have no difference between control and treatment of elicitor, that was friable until the end of treatment. The Calli with addition of elicitor Al3+ with 150 µM concentrations had significantly highest of fresh weight than control, they were over 0,463 g. The Content of daidzein and genistein on Phaseolus vulgaris L. had significantly increased in addition of Al3+ elicitor with 150 µM concentration. But, the content of daidzein and genistein on Phaseolus aureus Roxb. had significantly increased in addition of Al3+ elicitor with 300 µM concentration.&#x0D; &#x0D; Keywords: Phaseolus vulgaris L., Phaseolus aureus Roxb., elicitor Al3+, Daidzein, Genistein, Callus Growth.

Offset-free IMC with generalized disturbance models

Integral action is an essential component for offset-free response in many feedback control systems. Standard internal model control design rules ensure an offset-free response for open-loop stable plants. In this paper a linear feedback control structure is proposed that comprises an observer using an augmented plant model, state estimate feedback and disturbance estimate feedback. This allows internal model control design principles to be applied to unstable and marginally stable plants. Conditions are given for both nominal internal stability and offset-free action even in the case of plant-model mismatch. The Youla parameterization is recovered as a limiting case with reduced-order observers. The design is illustrated on a large scale non-square model with marginally stable dynamics.

Plant wax evidence for precipitation and vegetation change from a coastal sinkhole lake in the Bahamas spanning the last 3000 years

Plant wax hydrogen isotopic composition is commonly used to reconstruct the hydrogen isotopic composition of precipitation used by terrestrial vegetation. However, mangroves growing in coastal environments take up a mixture of freshwater and seawater. Biosynthetic fractionation (between source water and plant wax) differs between plant types and as a function of salinity, potentially complicating interpretations of past precipitation in coastal environments. In order to reconstruct Holocene hydrologic and ecologic changes archived within sediments from Blackwood Sinkhole on Abaco Island in The Bahamas, we adopt a multi-proxy approach using plant wax hydrogen isotopic composition (δ2H) to reconstruct paleohydrology, together with plant wax carbon isotopic composition (δ13Cwax), sterol biomarkers and pollen abundances to identify vegetation change. When pollen indicates a stable terrestrial plant community (2950–850 cal yrs BP), variations of δ2H values measured on the plant wax C28n-alkanoic acid are interpreted in terms of precipitation isotope (δ2Hprecip) changes, with 2H-depletion from 2950 to ∼2100 cal yrs BP and ∼1700 to 1000 cal yrs BP. However, interpretation is complicated at 850 cal yrs BP, when δ2H values decrease (−50‰) concurrent with increased Laguncularia racemosa (white mangrove) and Conocarpus erectus (buttonwood mangrove), and the mangrove-derived biomarker, taraxerol. We develop a pollen-based correction for mangrove inputs, yielding reconstructed precipitation isotope estimates (δ2Hprecip-corr). Low δ2Hprecip-corr values are synchronous with increased abundance of pine pollen, both of which may indicate wetter conditions from 850 cal yrs BP to present. This study provides additional evidence that mangroves can complicate hydrologic reconstructions from n-alkyl terrestrial plant wax biomarkers, and that such complication can be removed by pollen-based correction. After correcting for mangrove inputs, we obtain estimates of δ2Hprecip-corr from −33 to +25‰ throughout the last 2950 years, with uncertainties on the order of 10–20‰.

In vitro propagation of Salvia sclarea L. by meta-Topolin, and assessment of genetic stability and secondary metabolite profiling of micropropagated plants

The influence of meta-Topolin (mT) on micropropagation of Salvia sclarea L. (clary sage) was compared with N6-benzylaminopurine (BAP), kinetin (KIN) and thidiazuron (TDZ). mT was superior over all other cytokinin for not only shoot regeneration but also rooting. The highest shoot number (4.9 shoots/explants) was noted on nodal segments cultured on Murashige and Skoog (MS) medium including 2.0 mg/L mT and 0.2 mg/L indole-3-acetic acid (IAA). The regenerated shoots derived from mT including medium were rooted successfully on MS medium having 1.0 mg/L 1-naphthaleneacetic acid (NAA). Plantlets were successfully acclimatised (100%) and exhibited vigorous development in the greenhouse. The genetic stability of the propagated plants was assessed by start codon targeted polymorphism (SCoT) primers. A total variability of 3.13% was noted in the regenerants that revealed a high rated of genetic stability among the micropropagated plants. In addition, the secondary metabolite profiling of micropropagated plants was evaluated with gas chromatography-mass spectrometry (GC–MS). Although the hexane extracts of plants were dominated by n-alkanes, cytokinins added in the culture media altered their secondary metabolite contents. The protocol described here is a rapid and reliable micropropagation protocol for large-scale production of clonally stable clary sage plants for commercial purposes.

Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars.

This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars.

Stochastic Event-Based Sensor Schedules for Remote State Estimation in Cognitive Radio Sensor Networks

We consider the problem of communication allocation for remote state estimation in a cognitive radio sensor network~(CRSN). A sensor collects measurements of a physical plant, and transmits the data to a remote estimator as a secondary user (SU) in the shared network. The existence of the primal users (PUs) brings exogenous uncertainties into the transmission scheduling process, and how to design an event-based scheduling scheme considering these uncertainties has not been addressed in the literature. In this work, we start from the formulation of a discrete-time remote estimation process in the CRSN, and then analyze the hidden information contained in the absence of data transmission. In order to achieve a better tradeoff between estimation performance and communication consumption, we propose both open-loop and closed-loop schedules using the hidden information under a Bayesian setting. The open-loop schedule does not rely on any feedback signal but only works for stable plants. For unstable plants, a closed-loop schedule is designed based on feedback signals. The parameter design problems in both schedules are efficiently solved by convex programming. Numerical simulations are included to illustrate the theoretical results.