Mosquitoes have always been a human health threat; the major global health problems caused by them are malaria, dengue fever, yellow fever, and Zika as well as several other vector-borne outbreaks. The major problems in controlling these vectors borne diseases are related to resistance to eradication measures. Different classes of insecticides used for controlling public health have raised the concern of resistant problems with mosquitoes and environmental pollution caused by the control measures. Thus, a search for alternative natural compounds is necessary for solving the insecticidal resistance problem using pesticides in the larval stage of vector development as well as creating a chemical-free environment for a healthy society. Hence, the major focus of this study is to identify the larvicidal mechanisms, metabolite, antioxidants, and chemical compounds and elucidate their structures from C. ternatea flower and to test their efficacies against early 4th instar larvae of Aedes aegypti and Aedes albopictus. Clitoria ternatea flowers were collected from the garden of the Faculty of Medicine in International Quest University, Ipoh, Perak, and thence used for crude extraction. Further on, the metabolite test, antioxidant test, and chromatography techniques were conducted to identify the chemical composition of extracts and their chemical structures were identified using GCMS-QP2010 Ultra (Shimadzu). Next, the extracts were evaluated against the early 4th instar larvae of Aedes mosquito vectors following the WHO procedures for larval bioassays. The larvicidal activity of Clitoria ternatea flower extracts evidently affected the early 4th instar larvae of Aedes mosquito vectors. The highest larvicidal activity was observed against the early 4th instar larvae of Aedes aegypti with the LC50 and LC95 values of 1056 and 2491 mg/L, respectively. Meanwhile, the larvae bioassay test for Aedes albopictus recorded the LC50 and LC95 values of 1425 and 2753 mg/L. Moreover, the results for nontarget organism test on guppy fish, Poecilia reticulata, showed no mortalities with flower extracts at 2500 mg/L, hence posing no toxic effects on fish. In this study, we have found a total of 16 chemical compounds and 6 chemical compounds have been reported to possess direct insecticidal, larvicidal, and pupicidal effects. Six chemicals with insecticidal properties were found to be glycerin, 2-hydroxy-gamma-butyrolactone, neophytadiene, n-hexadecanoic acid, cis-vaccenic acid, and octadecanoic acid with a total of 28.7% efficacy. Clitoria ternatea flower extracts also showed different types of phenols such as anthocyanins, flavonoids, and tannins. Our findings showed that the crude extract of Clitoria ternatea flower bioactive molecules is effective and may be developed as biolarvicide for Aedes mosquito vector control. Furthermore, this study also provided a baseline understanding for future research work in the field of applications of Clitoria ternatea flower extracts for their long-term effects on human health such as a food additive, antioxidant, and cosmetic.

In germinating jack pine, changes in nitrogenous compounds were separated into two phases, the first, where through imbibition, seeds expanded but the prospective seedling remained enclosed by the haploid and nutritive female gametophyte (0 to 3 days), and second, when radicles emerged and only cotyledons remained in contact with the gametophyte (4 to 11 days).During imbibition, total soluble N in seeds dropped and the amino acid pool was dominated by high levels of free arginine. As levels of arginine N declined the greatest changes in percentage composition involved glutamic acid (gametophyte) and glutamine (embryo). Thereafter, arginine N accumulated. By 7 days, arginine N was recovered in seedlings primarily from cotyledons. High asparagine levels were observed in stems and roots as glutamine N in the emerging seedling declined.Protein reserves in the seedling were nearly depleted by 4 days. Total protein and, at later stages, the ratio of hexone bases to dicarboxylic acids was generally higher in the gametophyte than in the seedling. Soluble proteins of the embryo were separated into at least 18 bands by disc electrophoresis and contained peroxidase activity which increased strongly after the first week of germination. The increase of nine isoenzymes of peroxidase with mobilities towards the anode correlated with the histochemical localization of peroxidase at the emerging shoot and root tips and throughout the vascular tissues.

1971. The metabolism and subcellular organization of the jack pine embryo (Pinus banksiana) during germination. Can. J. Bot. 49: 927-938. Les graines de Pin grts (Pinus but~ksiat~a) imbibCes d'eau absorberent rapidement de I'oxygene et 95% d'entre elles germerent. A mesure que les cellules des cotyledons grossissaient et se divisaient, la redistribution des hydrates de carbone, protdines et acide~ nucleiques, sous observation cytochimique, correspondait a certains chanqements dans I'organisation subcellulaire.Chez !es cotyledons dormants, il n'y avait aucun chloroplaste dans les cellules; cependant les rberves en lipides, les protides et les arnyloplastes abondaient. Mais apres 97 h dans I'eau, les rCserves en nourriture furent entiirernent consurrees, le nouveau cytoplasme apparut et de nombreuses particules similipolyscrnes quitterent le noyau et se repandirent dans le cytoplasme. Correspondant au premier cycle mitotique, les proportions de nucleotides solubles dans I'acide, d'ARN et d'ADN augnlentirent pendant les 4 premiers jours de la germination. L'ARN etait concentre dans les phrag~nosomes et dans les plaques cell~~laires nouvellement formees. Les proteines acidiques, d'abord trouvees surtout dans les protides des cellules seches, se trouvaient maintenant dans les nucleoles. Les histones Ctaient surtout associes aux chromosomes, bien que les proteines de reserve, riches en arginine, fussent observees dans les protides des cellules dormantes. Les proplastides, qui tventuellement se developperent dans les chloroplastes demeuraient t o~~t pres du noyau jusqu'a I'utilisation quasiment totale des reserves cytoplasrniques.Au moment oh le cotyledon etait "sevre," le cytoplasme des cellules en pleine croissance contenait plus de mitochondries, aussi de I'endoplasme brut reticulaire, des ribosomes, des appareils de Golgi et des chloroplastes a au pluq quatre grains d'amidon. Dix A d0ui.e jours apres le debut de la germination, le cytoplasme apparut sur le pourtour des membranes cellulaires par suite, de I'accumulation d'eau dans une grande vacuole.Water imbibed by jack pine seeds induced rapid oxygen uptake and over 95% germination. As cells of cotyledons expanded and divided the redistribution of carbohydrates, proteins, and nucleic acids, followed cytochemically, corresponded to changes in subcellular organization.In dormant cotyledons, cells lacked chloroplasts and were packed with lipid reserves, protein bodies, and a~nyloplasts. After 96 h of imbibing water, storage materials were consumed as new cytoplasm appeared and numerous polysome-like particles extended from the n~tcleus into the cytoplasnl. Levels of acid-soluble n~~cleotides, RNA. and DNA increased up to the f o~~r t h day of germination coinciding with the completion of the first mitotic cycle. RNA was concentrated in phragmoso~nes and at the newly formed cell plates. Acidic proteins, found mainly in the protein bodies of dry cells, were now localized in nucleoli. Histones were associated mainly with chromosomes, although in dormant cells, reserve pr...

In hypocotyls of dry jack pine seeds, RNA was localized mainly in nucleoli and throughout nuclei. After 4 days of imbibition, RNA increased in the cytoplasm and was concentrated particularly in phragmosomes during the first wave of cell divisions. At 5-6 days, when radicles had emerged, uracil-5 or 6-3H was readily degraded to β-alanine via dihydrouracil and β-urcidopropionic acid. The remaining 10% of the radioactivity was recovered mainly as uracil from ribosomal RNA. Polyacrylamide gel electrophoresis was used to separate RNA into ribosomal and transfer RNA fractions. Within 90 min up to 70% of the tritium incorporated into RNA was recovered from 18 and 25 S ribosomal RNA (0.7 and 1.3 × 106 daltons). At the shoot apex, radioactivity per unit area was localized nearly 2-fold more in nuclei than in the cytoplasm and particularly in nuclei from the anneau initial (peripheral zone). At later exposure times to uracil, more radioactivity was detected in the cytoplasm and throughout the shod apex.

In germinating pine seedlings, acid soluble nucleotides, initially rich in AMP, accumulated, and were increasingly dominated by ATP. By 7 days, when hypocotyls were green but still dependent on the female gametophyte, the percentage of adenine nucleotides declined as other nucleotides increased. At 11 days, when gametophytic reserves were depleted, and seedlings were exposed to 32P- phosphoric acid, adenine nucleotides accounted for over 70% of the recovery of 32P from the nucleotide fraction.In seedlings, total RNA per unit weight increased to the 6th day, then levelled off. Comparison of electrophoretic mobilities of pine RNA on 2.5% polyacrylamide gels with ribosomal RNA from other sources revealed the following main types based on estimated molecular weights: 1.3, 1.1, 0.7, and 0.56 × 106 daltons, corresponding to 25S, 23S, 18S, and 16S, respectively. On 10% polyacrylamide gels, the mobility of low molecular weight pine RNA was similar to that of yeast tRNA. The 1.3 and 0.7 × 106 dalton fractions, representing ribosomal RNA, contributed 60–90% of the total RNA. During germination, the ratio of 1.3 to 0.7 × 106 RNA dropped from 2.4 to 1.7 and reciprocated the pattern for water content of the seedling.32P was incorporated mainly into the 25S and 18S RNA by 11-day-old seedlings, indicating that synthesis of ribosomes was a major event in growing seedlings. During germination, the synthesis of RNA corresponded to an increase of total RNA detected cytochemically and to an increase of new cytoplasm with more ribosomes, as observed previously with hypocotyl cells by electron microscopic procedures.