Computer-Based Tools Unmask Critical Mineral Nutrient Interactions in Hoagland Solution for Healthy Kiwiberry Plant Acclimatization

The aim of this study was to better understand the response of ex vitro acclimatized plants grown to a set of mineral nutrient combinations based on Hoagland solution. To reach that, two computer-based tools were used: the design of experiments (DOE) and a hybrid artificial intelligence technology that combines artificial neural networks with fuzzy logic. DOE was employed to create a five-dimensional IV-design space by categorizing all macroelements and one microelement (copper) of Hoagland mineral solution, reducing the experimental design space from 243 (35) to 19 treatments. Typical growth parameters included hardening efficiency (Hard), newly formed shoot length (SL), total leaf number (TLN), leaf chlorophyll content (LCC), and leaf area (LA). Moreover, three physiological disorders, namely, leaf necrosis (LN), leaf spot (LS), and curled leaf (CL), were evaluated for each treatment (mineral formulation). All the growth parameters plus LN were successfully modeled using neuro-fuzzy logic with a high train set R2 between experimental and predicted values (72.67 < R2 < 98.79). The model deciphered new insights using different sets of “IF–THEN” rules, pinpointing the positive role of Mg2+ and Ca2+ to improve Hard, SL, TLN, and LA and alleviate LN but with opposite influences on LCC. On the contrary, TLN and LCC were negatively affected by the addition of NO3– into the media, while NH4+ in complex interaction with Cu2+ or Mg2+ positively enhanced SL, TLN, LCC, and LA. In our opinion, the approach and results achieved in this work are extremely fruitful to understand the effect of Hoagland mineral nutrients on the healthy growth of ex vitro acclimatized plants, through identifying key factors, which favor growth and limit physiological abnormalities.

PENGARUH NAA DAN IBA TERHADAP PERAKARAN PURWOCENG (Pimpinella pruatjan Molk.) 77V VITRO

<p>ABSTRAK<br />Purwoceng (Pimpinella pruatjan Molk.) merupakan tanaman obat<br />langka yang cukup potensial untuk dikembangkan sebagai bahan baku<br />afrodisiak. Untuk mendukung budidaya tanaman ini diperlukan bahan<br />tanaman yang memadai. Perbanyakan in vitro purwoceng untuk<br />memperoleh bahan tanaman secara masal masih dibatasi oleh sulitnya<br />menginduksi akar, yang berakibat rendahnya keberhasilan aklimatisasi di<br />lapangan. Penelitian bertujuan untuk mendapatkan teknik induksi<br />perakaran dengan menggunakan dua jenis auksin (NAA dan IBA) pada<br />berbagai taraf konsentrasi yaitu 0; 0,1; 0,2; 0,4; 0,6; 0,8; 1,0; 1,5 dan 2,0<br />mg/1. Penelitian ini dilaksanakan di Laboratorium Kultur Jaringan, Balai<br />Penelitian Tanaman Rempah dan Obat, Bogor dari bulan Januari 2003<br />sarnpai dengan Februari 2004. Rancangan percobaan yang digunakan<br />adalah acak lengkap dengan tiga ulangan. Setiap ulangan terdiri dari tiga<br />tunas. Parameter yang diamati adalah jumlah akar, panjang akar dan<br />jumlah daun layu serta penampakan kultur secara visual. Hasil penelitian<br />menunjukkan bahwa auksin NAA nyata menghasilkan jumlah akar lebih<br />banyak dan lebih panjang dari IBA. Penggunaan NAA 0,8 g/1 merupakan<br />konsentrasi terbaik untuk induksi akar. Tidak ada perbedaan yang nyata<br />dari penggunaan NAA atapun IBAterhadap parameter jumlah daun layu.<br />Kata kunci : Purwoceng, Pimpinella pruatjan Molk, tanaman obat,<br />pengatur tuumbuh , NAA, IBA, induksi akar, in vitro, Bogor</p><p><br />ABSTRACT<br />Effect of NAA and IBA on root induction of pruatjan<br />(Pimpinella pruatjan Molk.) in vitro<br />Pruatjan (Pimpinella pruatjan Molk.) is one of endangered species<br />which is potential to be developed as aphrodisiac source. To support<br />pruatjan cultivation , it is needed to prepare the planting material. In vitro<br />propagation of pruatjan is hampered by the difficulty in inducing the<br />normal roots which affect the successful plant acclimatization. The<br />objective of this research was to obtain the root induction technique using<br />two kinds of auxin (NAA and IBA ) at several concentrations i.e : 0; 0.1;<br />0.2; 0.4; 0.6; 0.8; 1.0; 1.5 and 2.0 mg/1. This experiment was conducted<br />from January 2003 to February 2004 at the Tissue Culture Laboratory of<br />Indonesian Spices and Medicinal Crops Research Institute (ISMECRI) in<br />Bogor. Experiment was designed as a completely randomized design with<br />three replications. Each replication consisted of three shoots. The<br />parameters observed were number of roots, length of roots, number of<br />senessence leaves and culture performance. The result showed that NAA<br />produced the greatest and the longest roots compared to that of DBA. The<br />use of NAA 0,8 mg/1 performed the best treatment to induce roots. The<br />number of senessence leaves was neither affected by NAA nor IBA.<br />Key words: Pruatjan, Pimpinella pruatjan Molk., medicinal plant,<br />growth regulator, NAA, IBA, root induction, in vitro</p>

Somatic Embryogenesis from Mature Embryos of Olea europaea L. cv. ‘Galega Vulgar’ and Long-Term Management of Calli Morphogenic Capacity

Several olive cultivars, characterized by high-quality olive oil show agronomical issues such as excessive vigor, high susceptibility to biotic and abiotic stresses, and low propagation ability. They are strong candidates for breeding based on new technologies to improve their performance in a short period of time. For this reason, the first step is developing efficient somatic embryogenesis (SE) protocols. Somatic embryogenesis in olive is highly genotype-dependent for both adult tissues and mature embryos as initial explants, requiring the development of specific protocols for each genotype. Trials using cotyledons and radicles as initial explants, isolated from ripe seeds from the Portuguese olive cv. ‘Galega vulgar’, gave more than 95% calli development. Radicles proved to be the most responsive tissue for SE induction, with an average of 2 embryos per callus after callus transfer to expression medium, and 14 embryos per callus after subculture on the olive cyclic embryogenesis medium (ECO). Embryogenic competence could be recovered after several subcultures on ECO medium that maintained cyclic embryogenesis for an indeterminate period of time. Embryo conversion and plant acclimatization were also attained with high success rates. Media management for cyclic embryogenesis maintenance is of general importance for SE protocols in any olive genotype. Somatic embryogenesis was thus attained for the first time in embryo-derived explants of cv. ‘Galega vulgar’.

CtACO1 Overexpression Resulted in the Alteration of the Flavonoids Profile of Safflower

Background: Flavonoids with various structures play a vital role in plant acclimatization to varying environments as well as in plant growth, development, and reproduction. Exogenous applications of ethylene and 1-aminocyclopropane carboxylic acid (ACC), could affect the accumulation of flavonoids. Very few attempts have been made to investigate the effect of 1-aminocyclopropane carboxylic acid oxidase (ACO), a unique enzyme that catalyzes ACC to ethylene, on genes and metabolites in the flavonoid biosynthetic pathway. In this study, two ACOs in safflower (CtACOs) were cloned, and then transgenic safflower with overexpressed CtACO1 was generated through the Agrobacterium-mediated floral dipping method. Results: CtACO1 and CtACO2 were both characterized by the 2-oxoglutarate binding domain RxS and the ferrous iron binding site HxDxnH as ACOs from other plants. However, the transcript levels of CtACO1 in flowers at stages I, II, III, and IV were all higher than those of CtACO2. At the cellular level, by using electroporation transformation, CtACO1 was found to be localized at the cytomembrane in onion epidermal cells. CtACO1 overexpression had varying effects on genes involved in the ethylene and flavonoid biosynthetic pathways. The metabolites analysis showed that CtACO1 overexpression lines had a higher accumulation of quercetin and its glycosylated derivatives (quercetin 3-β-d-glucoside and rutin). In contrast, the accumulation of quinochalcones (hydroxysafflor yellow A and carthamin), kaempferol glycosylated derivatives (kaempferol-3-O-β-rutinoside and kaempferol-3-O-β-d-glucoside), apigenin, and luteolin in CtACO1 overexpression lines were decreased. Conclusion: This study confirmed the feasibility of applying the floral dipping method to safflower and showed a novel regulatory effect of CtACO1 in the flavonoid biosynthetic pathway. It provides hypothetical and practical groundwork for further research on regulating the overall metabolic flux of flavonoids in safflower, particularly hydroxysafflor yellow A and other quinochalcones, by using appropriate genetic engineering strategies.

Germination, in vitro Propagation and Acclimatization in Lavandula Angustifolia

This research focused on finding the best method for seed in vitro germination in Lavandula angustifolia and optimizing the medium for plant propagation. Seeds were sterilized and subjected to various treatments to break dormancy, then placed on half-strength MS (1/2MS) or distilled H2O + phytagel. Germination percentages were assessed and plantlets propagated on MS without growth regulators or with zeatin (0.5, 1, 2 mg/l), 1 mg/l BA + 0.5 mg/l IBA, 2 mg/l BA + 1 mg/l IBA or 3 mg/l BA + 1.5 mg/l IBA. After 8 weeks growth parameters were recorded and plants were acclimatized. Immersion in 20 mg/100 ml GA3 solution for 24 hours at 4°C was the most effective in breaking dormancy. Stratification at 4°C for 8 weeks and soaking in a solution of 0.5% H2O2 at 23°C for 24 hours also proved beneficial but to a smaller degree. Half-strength MS was the best germination medium. Shoot development was the highest in MS supplemented with zeatin (2 or 1 mg/l) while roots formed only in the control. Callus induction percentage was best in the presence of 3 mg/l BA + 1.5 mg/l IBA but decreasing concentrations increased callus weight. Plant acclimatization was more successful in moss:sand - 1:2 than in vermiculite:perlite:sand - 2:2:1.

Induksi Kalus dan Optimasi Regenerasi Empat Varietas Padi melalui Kultur In Vitro

<p class="p1">A study was conducted at the Tissue Culture Laboratory of ICABIOGRAD, Bogor, to obtain an optimum medium formulation for calli regenerations of for rice varities (Ciherang, Cisadane, IR64, and T-309). The research activities were done in five steps, i.e., callus induction, callus regeneration, shoot multiplication, root formation, and plant acclimatization. The type of explants used in the study was embriozygotic explants. Five media formulations were used for the callus induction, while four media formulations were used for the callus regeneration. The results showed that the best medium formulation for induction of callus formation was MS + 2,4-D 2 mg/l + casein hidrolisat 3 mg/l, while the best medium formulation for callus regeneration was MS + BA 3 mg/l + thidiazuron 0,1 mg/l.</p>

Anatomical traits of seven corn-inbred lines including two with gene brown midrib

Anatomical investigations of the stem in seven <em>Zea mays</em> L. Inbred lines were performed on specimens bred in the Experimental Institute of Breeding and Plant Acclimatization in Smolice. Two of the lines (bm1 and bm2) including the gene brown midrib were characterized by a higher digestability. The remaining five lines (S215, S335, 5336, S336A and S339) were selective inbred lines used as components in hybrid breeding at the Institute in Smolice. The investigated lines were compared in respect to 50 anatomical traits of the stem. The comparisons were performed by means of the Wrocław dendrite method. The lines formed three distinct groups according to the degree of similarity. The first group consisted of two lines with the gene brown midrib (bm1 and bm2), the second of four lines (5215, S336, S336A and S339), and the third of line S335. The inclusion of both the lines with gene bm into one group was based on similarity regarding the set of traits of parenchyma, particularly of the peripheral part of the stem, as well as metaxylem and metaphloem traits. However, these lines differed considerably in respect to epidermis traits. It was peculiar that the stomata of the <em>Amaryllis</em> type occurred in one of the lines (S339). Each line made a specific mosaic of traits. The sets of traits characterizing the particular lines were specific in such a degree that they could be used, like a fingerprint, for their identification.

Nemesia Root Hair Response to Paper Pulp Substrate for Micropropagation

Agar substrates forin vitroculture are well adapted to plant micropropagation, but not to plant rooting and acclimatization. Conversely, paper-pulp-based substrates appear as potentially well adapted forin vitroculture and functional root production. To reinforce this hypothesis, this study comparesin vitrodevelopment of nemesia on several substrates. Strong differences between nemesia roots growing in agar or in paper-pulp substrates were evidenced through scanning electron microscopy. Roots developed in agar have shorter hairs, larger rhizodermal cells, and less organized root caps than those growing on paper pulp. In conclusion, it should be noted that in this study,in vitromicroporous substrates such as paper pulp lead to the production of similar root hairs to those found in greenhouse peat substrates. Consequently, if agar could be used for micropropagation, rooting, and plant acclimatization, enhancement could be achieved if rooting stage was performed on micro-porous substrates such as paper pulp.