Stem anatomy and ontogeny of vascular variants in Ipomoea asarifolia (Desr.) Roem. & Schult. (Convolvulaceae)

Agarwoods such as Aquilaria spp. and Gyrinops spp. (Thymelaeaceae) produce interxylary phloem in their secondary xylem and intraxylary phloem at the periphery of the pith, facing the primary xylem. We studied young shoots of Aquilaria sinensis and characterized the development of its intraxylary phloem. It was initiated by the division of parenchyma cells localized in the outer parts of the ground meristem immediately following the maturation of first-formed primary xylem. Its nascent sieve plates bore donut-like structures, the individual pores of which were so small (less than 0.1 μm) that they were hardly visible under FE-SEM. Intraxylary phloem developed into mature tissue by means of the division and proliferation of parenchyma cells. During the shoots’ active growth period, the sieve pore sizes were 0.1–0.5 μm, with tubular elements passing through them. In the maturation stage, large clusters of sieve tubes continued to be differentiated in the intraxylary phloem. In the partial senescence stage observed in a three-centimeter-diameter branch, intraxylary phloem cells in the adaxial part became crushed, and sieve plates had pores over 1–2 μm in diameter without any callose deposition. Before and after the differentiation of interxylary phloem in the first and second internodes, callose staining detected more than twice as many sieve tubes in intraxylary phloem than in external phloem. However, after differentiation of interxylary phloem in the eleventh internode, more sieve tubes were found in interxylary phloem than in intraxylary and external phloem. This suggests that prior to the initiation of interxylary phloem intraxylary phloem acts as the principal phloem. After its differentiation, however, interxylary phloem takes over the role of principal phloem. Interxylary phloem thus acts as the predominant phloem in the translocation of photosynthates in Aquilaria sinensis.

Development of successive cambia and structure of secondary xylem in the stems and roots of Distimake tuberosus (Convolvulaceae)

Modification of external morphology and internal structure of plants is a key feature of their successful survival in extreme habitats. They adapt to arid habitats not only by modifying their leaves, but also show several modifications in their conducting system. Therefore, the present study is aimed to investigate the pattern of secondary growth in Leptadenia pyrotechnica (Forssk.) Decne., (Asclepiadaceae), one such species growing in Kachchh district, an arid region of Gujarat State. A single ring of vascular cambium, responsible for radial growth, divided bidirectionally and formed the secondary xylem centripetally and the phloem centrifugally. After a short period of secondary xylem differentiation, small arcs of cambium began to form secondary phloem centripetally instead of secondary xylem. After a short duration of such secondary phloem formation, these segments of cambium resumed their normal function to produce secondary xylem internally. Thus, the phloem strands became embedded within the secondary xylem and formed interxylary phloem islands. Such a recurrent behavior of the vascular cambium resulted in the formation of several patches of interxylary phloem islands. In thick stems the earlier formed non-conducting interxylary phloem showed heavy accumulation of callose on the sieve plates followed by their crushing in response to the addition of new sieve elements. Development of intraxylary phloem is also observed from the cells situated on the pith margin. As secondary growth progresses further, small arcs of internal cambium get initiated between the protoxylem and intraxylary phloem. In the secondary xylem, some of the vessels are exceptionally thick-walled, which may be associated with dry habitats in order to protect the vessel from collapsing during the dryer part of the year. The inter- and intraxylary phloem may also be an adaptive feature to prevent the sieve elements to become non-conducting during summer when the temperature is much higher.

Formation of inter-and intraxylary phloem in some species of Argyreia Lour. (Convolvulaceae)

Members of the Convolvulaceae are characterized by the climbing habit and occurrence of variant secondary growth. From a histological perspective, the genus Ipomoea L. is the most extensively studied, while other genera have been less studied. Here, stem anatomy of the least studied genus in the family, Hewittia Wight & Arn., represented by Hewittia malabarica (L.) Suresh was investigated using classical histological techniques. In both the samples collected from India and South Africa, stem thickness increased by developing different types of cambial variants such as: neo-formed vascular cylinders, parenchyma proliferation at the phloem wedges, ray-derived cambia from dilating phloem rays, internal cambium, intra- and interxylary phloem. Neo-formed vascular cylinders develop from the parenchyma cells external to the phloem as a meristemoid in thick stems and later in dilating ray cells. With the increase in stem diameter, cells of the phloem wedges showed proliferation by meristematic activity, which form a connection with the cortex by rupturing the primary tissue ring of eustele. Subsequently, development of cambium in phloem wedges and deposition of its derivatives increased the tangential width of rays. Mature thick stems (25–30 mm) give rise to a fissured stem. Intraxylary (internal) phloem development on the pith margin was observed from primary growth onwards and in thick stems secondary intraxylary phloem developed from the internal cambium. Internal cambium is functionally bidirectional and produces secondary xylem internally and secondary phloem externally. In all the samples, patches of unlignified parenchyma embedded within the secondary xylem dedifferentiate and mature into interxylary phloem with the increasing age. Development of cambial variant and structure of the secondary xylem is correlated with the functional significance of the climbing habit.

Interxylary phloem is here defined as strands or bands of phloem embedded within the secondary xylem of a stem or root of a plant that has a single vascular cambium. In this definition, interxylary phloem differs from intraxylary phloem, bicollateral bundles, pith bundles, and successive cambia. The inclusive but variously applied terms included phloem and internal phloem must be rejected. Histological aspects of interxylary phloem are reviewed and original data are presented. Topics covered include duration of interxylary phloem; relationship in abundance between sieve tubes in external phloem and interxylary phloem; distinctions between interxylary and intraxylary phloem; presence of parenchyma, fibers, and crystals in the interxylary phloem strands; development of cambia within interxylary phloem strands; three-dimensionalization and longevity of phloem, systematic distribution of interxylary phloem; physiological significance; and habital correlations. No single physiological phenomenon seems to explain all instances of interxylary phloem occurrence, but rapidity and volume of photosynthate transport seem implicated in most instances.

1. In Leptadenia spartium and L. reticulata there are three phloem regions: (a) the outer normal phloem, (b) the intraxylary or inner phloem, and (c) the interxylary phloem which forms inclusions in the wood. 2. The patches of intraxylary phloem arise from the pith cells, but in later stages even the xylem parenchyma cells adjacent to the pith take part in their formation. In old stems a cambium is differentiated on the outer faces of these phloem groups and produces some secondary phloem centripetally. 3. The interxylary phloem, present in the stem, becomes differentiated from groups of thin-walled cells produced centripetally by the cambium. Later the cambium resumes its normal activity with the result that the phloem groups become embedded in the secondary xylem. 4. Owing to an enlargement of the cells in the island and the fact that it is surrounded on all sides by the woody cells of the xylem, there is often a compression and crushing of the phloem tissues in its centre. 5. A weak secondary cambium has occasionally been observed to differentiate on one or more than one side of some of the older phloem islands. 6. A comparison with Strychnos nux-vomica shows that in the latter the islands are always produced centrifugally from the cambium and later become embedded due to the formation of a complementary cambial segment on the outer side, whereas in Leptadenia it is the same cambium which produces both the secondary xylem as well as the phloem on its inner side.

Stem flattening in Rhynchosia pyramidalis (Fabaceae) is achieved by the development of crescent-shaped successive cambia on two opposite sides of the stem (referred hereafter as distal side). Other lateral sides of the stem (adjacent to supporting host and its opposite side, referred as proximal sides) usually possess single cambium. In the young stems, parenchymatous cells located outside to protophloem of distal side dedifferentiate and develop small segments of cambium. Concomitant to bidirectional differentiation of the secondary xylem and phloem, these newly developed cambial segments also extend in tangential directions. Differential activity of newly developed crescent-shaped cambial segments deposits more secondary xylem at median position as compared to their terminal ends of the stem on distal side; consequently, it pushes the cambial segment outside, thus resulting in crescent-shaped arcs of the cambia only on two opposite sides. After the production of 1–2 mm of secondary xylem, they cease to divide and new segments of cambial arc develop on the same side in a similar fashion. Such repeated behaviour of successive cambia development consequently leads to the formation of tangentially flat stems. The secondary xylem is diffusely porous with indistinct growth rings and is composed of vessels (wide and narrow), fibres, axial ray parenchyma cells, while phloem consisted of sieve elements, companion cells, axial and ray parenchyma. Rays in both xylem and phloem are uni- to multiseriate and heterocellular. The structure of secondary xylem and development of successive cambia is correlated with climbing habit.

This study examined the formation of successive rings of cambia in Rivea hypocriteriformis Choisy (Convolvulaceae). The mature stem is composed of four to five rings of xylem alternating with phloem. Successive cambia originate as smaller and larger segments; union and anastomosing of small cambial segments often leads to the formation of discontinuous rings. In the initial stage of growth, several vascular bundles interconnect to form the first ring of vascular cambium. The cambium remains functional for one complete season and becomes dormant during summer; a new ring of cambium is completed prior to the subsequent monsoon season and sprouting of new leaves. Successive cambia are initiated from the pericyclic parenchyma situated three to four cell layers outside of the protophloem. Functionally, all the successive cambia are bidirectional and produce secondary xylem centripetally and phloem centrifugally. The secondary xylem is diffuse-porous, with indistinct growth rings and consisting of wide fibriform vessels, fibre tracheids, and axial and ray parenchyma cells. The xylem rays are uni- to multiseriate and heterocellular. The multiseriate rays contain lignified marginal ray cells and thin-walled, unlignified central cells. The central ray cells also show accumulations of starch and druses. Discrete strands of intraxylary phloem occur at the periphery of the pith, and additional intraxylary phloem develops from adjacent cells as secondary growth progresses. Earlier-formed phloem shows heavy accumulation of callose, followed by its compaction. The development of successive cambia is correlated with extension growth and with the phenology of the plant.

Summary The structure of axial and radial or ray sieve tubes was examined by fluorescence microscopy and SEM. Two types of sieve areas were found on the lateral walls of axial and radial sieve tubes: one consists of a cluster of larger, more highly differentiated sieve pores, corresponding to sieve plates, and the other consists of scattered or clustered minute sieve pores, arranged in a linear array in the latter along the cell axis, covered by slime bodies, making a new structure complex associated with nutrient pathways. Migration of cytoplasmic slime bodies in axial and radial sieve tubes was observed to take place by changing shape into filamentous or tubular slimes and passing through the sieve pores, depending on the size of the pores at the xylem differentiation stage. Axial sieve tubes in the interxylary phloem were observed to be arranged axially in dense bundles rather than sparsely scattered. Rarely, it was observed that axial sieve tubes themselves branched off and merged with other axial sieve tubes without being bridged by radial sieve tubes in the ray tissue. Parenchyma cells and ray tissue in the interxylary phloem were unlignified in all the secondary xylem from the 12-year-old tree of Aquilaria sinensis. The division of ray parenchyma cells was often observed in the mature secondary xylem. Furthermore, two types of radial sieve tubes at different stages of differentiation were observed in the same ray tissue, suggesting that at least younger radial sieve tubes differentiate from ray parenchyma cells in the secondary xylem. Based on the positioning of radial sieve tubes and axial sieve tubes in the secondary xylem, three types of connections were identified; (1) radial sieve tube strand in interxylary phloem connects to axial sieve tube strand; (2) radial sieve tube strand bridges two interxylary phloems; (3) radial sieve tube strand anastomoses with interxylary phloem and connects to axial sieve tube strand. The highest frequency of (1) among the three types suggests that it is crucial for the functioning of the phloem network and that radial sieve tubes and axial sieve tubes located inside the interxylary phloem are connected to establish longitudinal transport channels.

Structure of secondary xylem and pattern of secondary thickening in climbing species are different from those in self-supporting plants. In many climbing species, stem diameter increases by forming more than one ring of cambium (referred to as multiple/successive cambia), while their secondary xylem usually contains abundant parenchyma, large vessels and wide rays. In beach morning glory (Ipomoea pes-caprae (L.) R. Br., Convolvulaceae), stem thickness increases by forming multiple rings of cambia. After a short period of normal secondary growth, the first successive cambium ensues from the pericyclic parenchyma. Thereafter, subsequent cambial rings originate from parenchyma cells produced initially by the previous cambium. In stems that are 15–20 mm thick, parenchymal cells produced by the initial activity of the previous cambia become meristematic and form small arcs of functionally inverse cambia that produce secondary xylem centrifugally and secondary phloem centripetally. Unequal production of secondary xylem by these cambia gives the stem various shapes other than cylindrical. Besides successive cambia, some cambial variants also develop in the stem which are: (1) irregularly distributed patches of thin-walled xylem parenchyma becoming meristematic and differentiating into interxylary phloem islands; (2) xylem ray cells acquiring meristematic character and behaving like cambium (referred to as ray cambium); and (3) in thick stems, internal cambium deriving from marginal pith cells, which are functionally bidirectional and producing secondary xylem centripetally and phloem centrifugally. Structure and development of successive cambia, ray cambia and internal cambium are discussed here.

Background – Population growth of lianas in the tropical forest is credited to their ability of CO2 sequestration and efficiency of the narrow stems to supply water required for the amount of foliage it bears. Turbina corymbosa (L.) Raf. (Convolvulaceae Juss.) is one of the fast-growing invasive species of scrambling woody lianas. It covers trees entirely within a short period to compete with above-ground resources (particularly sunlight). However, no information is available on how it manages to cope up with an increasing demand of water supply and mineral nutrients. What are the structural and developmental patterns adapted by this species to expand the stem diameter for efficient supply of below-ground resources? Therefore, our aim was to investigate the secondary growth patterns and structure of secondary xylem and phloem in T. corymbosa.Methods – Several samples of the stem with various diameters were studied using a histological method. Morphological and anatomical analyses were carried out using light microscopy.Key results – With the initiation of secondary growth, stems lose their circular outline rapidly due to unequal deposition of secondary xylem and formation of successive cambia. New successive cambia initiate from parenchymatous cells as small crescent-shaped fragments on asymmetric/opposite sides and result in a different stem conformation. Though several segments of successive cambia are formed, very few stem samples form complete cambium rings. The secondary xylem formed by successive cambia is diffuse porous with indistinct growth rings and is composed of both wide and narrow (fibriform) vessels, tracheids, fibres, axial and ray parenchyma cells. The secondary phloem consists of sieve tube elements, companion cells, axial and ray parenchyma cells. In fully grown plants, cambial action (internal cambium) occurrs between the intraxylary phloem and protoxylem and produces secondary xylem and phloem near the pith region.Conclusion – Structural alterations and unequal deposition of conducting elements, occurrence of intraxylary phloem and flattening of the stem are suggested to facilitate rapid growth of the plants by providing required minerals and nutrients. Internal cambium formed at the periphery of the pith is bidirectional and produces secondary xylem externally and intraxylary phloem internally. Continued development of intraxylary phloem from the internal cambium provides an additional path for rapid and safe translocation of photosynthates.

Quantitative and qualitative features of wood and stem anatomy are presented for 44 collections of 16 genera and 35 species ofConvolvulaceae. Markedly furrowed xylem characterizes the genera of tribe Cresseae. Successive cambia occur in 11 of the genera studied. Large patches of axial parenchyma occur in many of these; only in one species was interxylary phloem (formed internally by the cambium) observed in the parenchyma patches. Intraxylary phloem at the periphery of the pith is universal in Convolvulaceae, but newly reported is the fact that in many species, cambial activity adds secondary phloem to the intraxylary phloem strands. These cambia were also observed to add limited amounts of secondary xylem externally in Ericybe and Operculina. Fragmentation of the vascular cylinder by growth from the inner cambia leads to initiation of radially oriented cambia (ray cambia) along the ray zone where fracture occurs. Three new types of vessel restriction patterns (nonrandom distribution of vessels in wood or absence of vessels within some parts of the secondary xylem) are reported for the family (and dicotyledons). Rays are predominantly uniseriate; ray histology and quantitative vessel features show little change ontogenetically in species with successive cambia, suggesting that this cambial mode deters the kinds of progressive changes that occur in dicotyledons with a single cambium. Vessels are much wider in lianoid Convolvulaceae than in shrubby species; the lianoid species of Ipomoea have vessels twice the diameter of those in arborescent species of Ipomoea. Conjunctive parenchyma may serve for water storage in I. arborescens and other species, but this tissue and abundance of axial parenchyma in lianas might also aid flexibility and damage resistance. Septate fiber-tracheids and septate libriform fibers occur in a few species and represent tracheid dimorphism. Occurrence of tracheids together with fibriform vessel elements in woods of many Convolvulaceae suggests relationship of Convolvulaceae to Polemoniaceae and Hydrophyllaceae; intraxylary phloem and other wood features suggest relationship between Convolvulaceae and Solanaceae.

Stems of Ipomoea obscura Ker Gawl., increase in thickness by forming multiple rings of cambia. Stems 5-6 mm thick produce parenchymatous derivatives which divide repeatedly to form small arcs of cambium. Several such small arcs initiate simultaneously and form a ring of small cambial arcs. After the formation of a few xylem and phloem elements, all these arcs are interconnected by transdifferentiation of parenchyma cells present between the cambial arcs and constitute a complete cambial cylinder. This newly formed cambium is functionally bidirectional: earlier- formed arcs produce xylem centripetally and phloem centrifugally, while later-formed segments exclusively produce thin-walled parenchyma cells on either side. Young stems are circular in cross section but as stem thickness increases they become oval to elliptic or lobed and dumbbell-shaped. Xylem rays are mostly uni- or biseriate and thin-walled, but multiseriate rays characteristic for a climbing habit are observed occasionally. In thick stems, the marginal ray parenchyma in most of the samples becomes meristematic and develops ray cambia which exclusively produce sieve elements. Similarly, parenchyma cells produced from later-formed cambial segments give rise to several irregularly oriented vascular bundles. The secondary xylem is diffuse porous, with indistinct growth rings and is composed of fibriform and wider vessels, fibres, and axial and ray parenchyma cells, while phloem consists of sieve elements, companion cells, and axial and ray parenchyma cells.

Phloem is one of the vital tissues of the vascular system that plays a crucial role in the conduction of photosynthates. In vascular plants, it develops external to the vascular cambium but in a small fraction of eudicots (formerly known as dicots), it occurs within (interxylary) and inside (intraxylary) the secondary xylem. Ontogenetically, it is classified as Strychnos, Combretum, Azima, and Calycopteris types. In all four cases, phloem islands remain enclosed within the secondary xylem but each has unique origins. Similarly, the deposition of the phloem at the pith margin is common in several plants. It develops from procambial derivatives or adjacent pith cells or by initiating an intraxylary phloem cambium. Functionally, this cambium can produce only phloem or both secondary xylem and phloem. In some instances, the deposition of the secondary xylem and phloem in the same direction has also been documented. Some experimental evidence is available on the role of phloem but is it applicable to inter- and intraxylary phloem? The presence of inter- and intraxylary phloem is attributed to a defence mechanism against insects or plants that show sudden and enormous flowering or it can correlate with high temperatures or an unconducive climate in a desert region where sieve tube elements have become nonfunctional due to high temperatures. The present review is an attempt to analyse the role of interxylary and intraxylary phloem.