Enraizamento de plantas cultivadas in vitro

The adventitious rooting process of in vitro cultured plantlets is a technique that has been employed for the vegetative propagation of a significant number of native and exotic species. Many factors are associated with the rooting stage influencing positive and/or negatively the establishment of micropropagation protocols. The objective of this work was a literature review of the main inherent factors concerning in vitro rooting process including the correlation among others the endogenous and exogenous auxins levels, juvenility, genotype, mineral nutrition, culture medium conditions, addition of growth regulators and other substances as phenolic compounds and active coal besides growth environmental conditions of in vitro cultures. Although the complete elucidation of all processes involved with rooting of in vitro cultured plants has not been achieved so far, a comprehensive study of the main factors that interfere on rooting is fundamental for the establishment of new researches that might contribute for the rooting of economically important plants.

Essential oil composition of Aloysia triphylla (L'Herit) Britton leaves cultivated in Botucatu, São Paulo, Brazil

The family Verbenaceae comprises about 175 genera and 2300 species, distributed in tropics and subtropics, mainly in temperate zone of southern hemisphere. The lemon verbena (Aloysia triphylla (L'Herit) Britton) is a perennial, bushy plant originally from South America. The essential oil of this plant is used in pharmaceutical, cosmetic and perfumery industry. Therapeutic properties include febrifuge, sedative, stomachical, diuretic, and antispasmodic activities. The present work aimed to identify the chemical composition of essential oil of Aloysia triphylla leaves. The study was done in Lageado Experimental Farm of the Department of Plant Production-Horticulture, Agronomical Sciences College, São Paulo State University Campus of Botucatu. Leaves of lemon verbena from Medicinal and Aromatic Plant Garden, were collected in the end of winter (September/2001). The essential oil was extracted by hydrodistillation, in Clevenger apparatus. 100 g of leaves were used in each extraction. Four extractions were performed during three hours. The essential oils of the leaves were analyzed in Gas Chromatography Mass spectrometry (CG-MS, Shimadzu, QP-5000), equipped with capillary column DB-5 (30 m × 0,25 mm × 0,25 mm), split 1/35, injector for 220 C°, detector for 230 C°, dragged by gas He (1,0 mL/min), with programmed temperature for 60 C° to 240 C°, 3 C°/min. The identification of the substances was held by comparison of their mass spectra with data of the CG-MS (Nist 62 lib), literature references and retention index of Kovats. The main constituents of essential oils were geranial (29.54 %), neral (27.01 %), limonene (15.93 %), geranyl acetate (4.0 %) and geraniol (3.96 %). This species possesses high quantity of monoterpenes and low quantity of sesquiterpenes.

Influence of IBA and boric acid on rooting of stem-cuttings of Aloysia triphylla (L'Hérit) Britton

Aloysia triphylla (L'Hérit) Britton is a perennial and bushy plant, with simple, entire, lanceolate and whorl shaped leaves and originally from South America. It is used as medicinal plant in Brazil with stomatic and sedative properties. The employment of stem cuttings for propagation of pre-selected plants, acquired great importance, because it eliminates the juvenile phase of seedlings, which can be produced in a shorter period of time. The rooting of stem cuttings is stimulated by auxin and, boric acid supply is essential for growth and development of initial rootlets. This micronutrient is required 48 hours after plant segments have been placed into auxin solution and it can be supplied any time, including the seedling growth period. The experiment was carried out in the Department of Plant Production, UNESP-Botucatu-SP-Brazil, with stem-cuttings of Aloysia triphylla (L'Hérit) Britton, Verbenaceae obtained from the Medicinal and Aromatic Plant Garden. The aim of the work was to verify the influence of growth regulators and boric acid on stem cutting rooting of this species. The 15 cm-stem cuttings, without leaves, were submerged during 24 hs in the following solutions: water; 150 mg.L-1 of IBA; 150 mg.L-1 of IBA+ Boric acid; 250 mg.L-1 of IBA; 250 mg.L-1 of IBA + Boric acid. The statistical design was entirely randomized with 5 treatments and 3 replications, totalizing 15 plots with 10 stem cuttings each. They were planted on propylene trays with vermiculite and kept under spraying condition during twenty five days. The best results were observed in treatment 250 mg.L -1 of IBA+ Boric acid on number of roots, length of roots, rooting percentage, fresh and dry weight of leaves when compared with all other treatments. We can conclude that this treatment is the most suitable for stem cutting rooting of this species.

Arisaema triphylla

R. Anheisser

Ruta graveolens - Adenophora suaveolens - Lysimachia nummularia

AR

Life history and host range of Alagoasa extrema (Coleoptera: Chrysomelidae: Alticinae), a potential biological control agent of Lantana spp. (Verbenaceae) in Australia

The life history and host range of the lantana beetle, Alagoasa extrema, a potential biocontrol agent for Lantana spp. were investigated in a quarantine unit at the Alan Fletcher Research Station, Brisbane, Australia. Adults feed on leaves and females lay batches of about 17 eggs on the soil surface around the stems of plants. The eggs take 16 days to hatch and newly emerged larvae move up the stem to feed on young leaves. Larvae feed for about 23 days and there are three instars. There is a prepupal non-feeding stage that lasts about 12 days and the pupal stage, which occurs in a cocoon in the soil, lasts 16 days. Teneral adults remain in the cocoon for 3 days to harden prior to emergence. Males live for about 151 days while females live for about 127 days. The pre-oviposition period is 19 days. In no-choice larval feeding trials, nine plant species, representing three families, supported development to adult. Three species, Aloysia triphylla, Citharexylum spinosum and Pandorea pandorana were able to support at least two successive generations. These results confirm those reported in South Africa and suggest that A. extrema is not sufficiently specific for release in Australia. Furthermore, it is not recommended for release in any other country which is considering biological control of lantana.

Using IRAP markers for analysis of genetic variability in populations of resource and rare species of plants

Species specific LTR retrotransposons were first cloned in five rare relic species of drug plants located in the Perm’ region. Sequences of LTR retrotransposons were used for PCR analysis based on amplification of repeated sequences from LTR or other sites of retrotransposons (IRAP). Genetic diversity was studied in six populations of rare relic species of plants Adonis vernalis L. by means of the IRAP method; 125 polymorphic IRAP markers were analyzed. Parameters for DNA polymorphism and genetic diversity of A. vernalis populations were determined.

Using IRAP markers for analysis of genetic variability in populations of resource and rare species of plants

Species specific LTR retrotransposons were first cloned in five rare relic species of drug plants located in the Perm’ region. Sequences of LTR retrotransposons were used for PCR analysis based on amplification of repeated sequences from LTR or other sites of retrotransposons (IRAP). Genetic diversity was studied in six populations of rare relic species of plants Adonis vernalis L. by means of the IRAP method; 125 polymorphic IRAP markers were analyzed. Parameters for DNA polymorphism and genetic diversity of A. vernalis populations were determined.

狭叶沙参复合体（桔梗科沙参属）的物种生物学研究

此复合体原先方7个种，1983年被处理为3个种。本文在黑龙江、吉林、辽宁、河北、山西、山东、江苏、内蒙古对本复合体进行了居群取样，共46个居群。其内进行了引种、栽培实验。生物学特性观察、杂交、染色体观察，同工酶比较，以及用生物统计学进行了性状分析、数量分类学分析、茎解剖和芯粉的观察。 此复合体主要进行异花授粉（因为雄蕊先熟)、没发现无融合生殖。实生苗当年只长基生叶，第二年才抽茎开花。 2、此复合体酯酶酶谱的变异范围相当宽，有时。同一居群内的变异反而比居群间的还要大。因此酶谱在本复合体内的分类价值不大。 3、通过对32个居群进行染色体记数，友现辽东、辽西(绝大部分地区)、山西霍县的居群为二倍体（2n=34），另外还在锦州发现有几个个体2n = 36。其它地方的居群都力四倍体，2n=68、居群间的染色体形态相当一致。 4、通过对移栽自黑龙江口(A•gmelinii的二个居群)、北京(A•polyantha)，山西东部(A•gmelinii和A•polyantha)，锦州的二倍体共9个居群进了杂交试验，发现四倍体间能得到一些F1种子、而辽西二倍体与其它类群间却得不到F1种子。 5、性状分析，茎解剖和数量分类研究发现花的大小和形状、果实的大小和齿的有无、叶的大小和形状（长/宽），往头与花冠相对长度和种子形状等都有一定的分化，尤其是髓锥管束的有无等。通过分析可以分辨出3个二倍体和5个四倍体宗。在 3个二倍体宗中，辽西西部宗(A•polyantha var•ootra ta)与其它两个在形态上已经分化得相当分明，无疑可以作种处理，A•contracta(Kitag) Qu et Hang，另二个（分别在辽东和山西霍县）在形态上也已分化得相当明显，达到了种水平，A•polyantha和A•gme1ireii在5个四倍体宗中，山西东北部和河北的西北部高海拔分布的一个宗，由于它的花萼裂片具齿且狭长，花冠狭长而大、果实大，茎常中空等特征，仍然保留种的地位（A•elata)。另外分布于山西高海拔(包括四倍和二倍体）以及内蒙、吉林、黑龙江…三个宗，虽然有分化，但不明显，尤其是它们都无髓锥管束或仅少数个体有的特征，本文把它们处理为同一种(A•gmelinii)的3个不同三种。最后一个是分布于辽宁最西部、河北、山东、江苏北部、安徽、河南、陕西和甘肃的四倍体宗、它在形态上与辽东的二倍体相似、故作为A•polyantha的两个亚种处理，前者为A•polyantna sspscabricalyx后者为A•P•ssp polyantha： 可以推测过去此复合体的分布范围相当宽，后来逐渐分化成三个二倍宗体，由辽东的A•polyantha ssplyantha(2x)衍生的四倍体，逐渐向西迁移，山西霍县的二倍体产生的四倍体逐渐向北迁移直至东西伯利亚东南部和远东，最后分化成三个亚种。 本文采用杂交指数法，对山西东部和河北西部的A•gme1inii和A•polyantha居群间的杂交性近行了研究，认为两个种在四倍体水平的杂交可能是这两个种间性状在该地区彼此过渡的主要原因。

泡沙参复合体的物种生物学研究

泡沙参复合体属沙参属组，原有6个种，从东北连续分布到西南，本研究从辽宁、内蒙古、河北、河南、山西、陕西、甘肃和四川等9个省、市、自治区，采集了22个居群（包括复合体原有的6个种）及筒花组的对照种(A.stcnanthina)一个居群．通过野外和移栽后的对比观测，播种栽培，染色体观察，杂交试验和子代测定，同工酶电泳，并结合多变量统计分析，对复合体从不同水平和角度进行了研究． 1．本复合体多年生，实生苗当年只长基生叶，第二年才抽茎开花，个体花朵很多，因而花期很长（可达2个月）．野生个体的结实量很大，种子萌发率接近100%．雌雄异熟，以异花授粉为主．自交（同株异花授粉）亲和，但结实量和萌发率均有所下降．等位酶估测的平均异交率(t)为0.77．末发现无融合生殖现象．全部居群的染色体均为2n= 34．末发现倍性和基数的变化． 2．同一个体的野生及栽培观测表明，根、茎、花序分枝等都是环境可塑性很高的性状，而叶片形态、花萼裂片长则有明显的发育可塑性，茎叶被毛、花部、果实和种子性状的可塑性较小. 3．针对在以往复合体类群划分中起鉴别作用的叶型、叶缘锯齿、茎叶被毛等几个性状，进行了大量的子代测定、杂交试验和天然居群的分析，结果表明，叶型、叶缘锯齿数目和大小、茎叶被毛等都是相关性很强的一组性状，但它们在天然居群中连续变化，是受多基因决定的数量性状．根据这些性状所划分的几个“种”，不过是连续变异的极端个体，极端个体（“种”）的自由授粉子代会分离出各种性状状态的个体，包括变异另一极端（另一个“种”）的个体，极端个体间的杂交进一步证实了这些数量性状的变异．据此，并结合地理分布，将A.bockiana和A.polydentata归并在A.potaninii之中：将A.biformifolia归并在A.wawreana之中．并证明A.wawreanavar. lanceifolia并不存在． 4．对20个居群34个形态性状的统计学和主成分分析表明，不同性状的系统学意义不一样，具有较大分类价值的性状是那些可塑性低、群内变异小的花部、果实、种子和部分营养性状.根据性状变异式样的分析和20个居群二套性状的聚类结果，可将复合体分为两个区别明显的类群．其中A.lobophylla形态十分独特，仅分布在四川西北部．另一个广布类群，在一些性状上体现出由西南(原A.potaninii)至东北(原A.wawreana)的地理变异式样，但性状在山西、陕西一带有过渡，是两个分化不充分的类群． 5．对复合体内8个居群及对照种居群的杂交试验表明，根据形态性状分析所归并的三个“种”与复合体其它类群间没有任何育性障碍，证实了归并的合理性。与此同时，发现根据形态变异式样所划分出的两个分化明显的类群(A.lobophylla与A.wawrcana和A.potaninii)间有很大的生殖隔离，不仅杂交结实量很低，杂交种子萌发率均小于10%．说明它们之间的亲缘关系较远，而A.lobophylla与筒花组的A.stenauthina间却有很高的杂交亲和性，平均结实量为53，与同种居群内的结实量相当;杂交种子萌发率达65%左右．只稍低于两个种各自居群内杂交的种子萌发率．因此，根据形态学，生境特点和杂交试验，建议将A.lobophylla放在远离本复合体的筒花组内． 6．采用控制杂交等方法确定了8个酶系统共10个等位酶位点的遗传基础．采用等位酶分析技术对复合体14个居群及一个对照种居群进行了测定．结果表明，本复合体植物的遗传变异水平很高，多态位点百分率(P)为79.2．等位基因平均数(A)为2.35，平均期望杂合度(He)为0.284．居群处于中等分化程度.总的等位酶变异中，有31%来自于居群间(FST=0.311)．而其余69%左右存在于居群内．相比之下，四川西北的特有种A.lobophylla遗传变异水平稍低(P=70.0，A＝2.0，He=0.211)，尤其是居群中的杂合体比例明显偏离理论期望比例，异交率估计值(t)为0.535. 7．本复合体丰富的遗传变异和高度的环境可塑性使其适应性强，分布范围广．物种形成的方式基本上是地理式的，南部居群和北部居群是两个分化不完全的地理宗，并仍处在强烈的分化之中．对本复合体、A.lobophylla和对照种A.stenanthina形态学、杂交及等位酶分析表明，沙参属起源较晚、进化速度较快.推测在其起源早期，历了一段快速的适应性辐射阶段，随后则以地理式物种形成过程为主，逐渐发展为当今这样一个中等大小的属． 8．通过上述各方面的研究，最后将本复合体原有的6种l变种归并为两个种，即A.potaninii Korsh.和A.lobophylla Hong．后者实际上不属于本复合体，而是另一个组－－筒花组的成员;而前者包含两个亚种：ssp.potaninii和ssp.wawreana Ge et Hong．