University of the Philippines Baguio

College of Science
Department of Biology



FOUR SUCCULENT FAMILIES AND 40 MILLION YEARS OF EVOLUTION AND ADAPTATION
TO XERIC ENVIRONMENTS: WHAT CAN STEM AND LEAF ANATOMICAL CHARACTERS TELL
US ABOUT THEIR PHYLOGENY
James V. Landrum










Ma. Rica Paulene B. Marquez
2012-68814
Bot 109 Pqr




I. SUMMARY
INTRODUCTION
Over the last 40 million years, four families of the Order Caryophyllales (Aizoaceae,
Cactaceae, Didiereaceae, and Portulaceae) have been particularly successful at adapting to xeric
environments, particularly over the last five million years as modern continental positions and
climate were stabilizing. The modifications of anatomy, morphology, and reproductive
strategies are the cause of the evolution and adaptation of the four families to xeric
environment which are not present in many other dicot families.
The study focuses on an evolutionary examination of stem and leaf anatomical
adaptations, and an attempt to link paleoclimatology to the evolution of these traits. The
primary hypothesis is that aridity islands formed in the ancestral areas of these families as
modern continental position and climates began to stabilize leading to the application of the
principles of island biogeography, such as accelerated evolution and founder effect.
METHODOLOGY
Twenty two species from six families were examined for the study: Achatocarpus gracilis
(Achatocarpaceae), Aptenia cordofolia (Aizoaceae), Galenia pubescens (Aizoaceae), Lampranthus
comptonii (Aizoaceae), Mesembryanthemum crystallinum (Aizoaceae), Sesuvium portulacastrum
(Aizoaceae), Anredera vesicaria (Basellaceae), Tournonia hookeria (Basellaceae), Ullucus
aborigineus (Basellaceae), Pereskia diaz-romeroana (Cactaceae), Alluaudia humberti
(Didiereaceae), Didierea trollii (Didiereaceae), Anacampseros arachnoides (Portulaceae),
Ceraria fruticolosa (Portulaceae), Cistanthe grandiflora (Portulaceae), Grahamia bracteata
(Portulaceae), Portulaca oleracea (Portulaceae), Portulacaria aria (Portulaceae), Talinaria
palmeri (Portulaceae), Talinella pachypoda (Portulaceae), Talinopsis frutiscens (Portulaceae),
Talinum paniculatum (Portulaceae). Plants used were either grown from field-collected seed or
were purchased as live plants. Living specimens were fixed with Navashins solution then
embedded in Paraplast Plus. Slides were stained using safranin and fast green dyes. Herbarium
materials were processed using a rehydration protocol and then process as above.
The stem and leaf anatomical characters were examined for the 22 included taxa. Only
taxa with both stem and leaf samples were used in order to prevent missing data. Most
characters were coded as binary; however, some characters were coded as multi-state. All
characters were unordered and unweighted except for the presence of anomalous secondary
growth and the type of photosynthesis.
A preliminary survey of potential characters yielded 51 stem and leaf characters found
to be informative: (1) stem epidermis, (2) stem epidermal projections, (3) stem epidermal
crystals, (4)stem epidermal cuticular waxes, (5) stem stomata, (6) stem phellem, (7) stem
hypodermis, (8) stem hypodermal crystals, (9) stem hypodermal collenchyma, (10) stem
hypodermal sclerenchyma, (11)cortical sclerenchyma, (12) cortical crystals, (13) cortical
mucilage, (14) cortical collenchyma, (15)cortical ducts, (16) phloem fibers, (17) ray
sclerenchyma, (18) ray collenchyma, (19) ray crystals, (20) ray wide-band tracheids, (21)
secondary xylem annual/helical pitting, (22) secondary xylem scalariform pitting, (23)
secondary xylem reticulate pitting, (24) secondary xylem circular bordered pitting,
(25)secondary xylem fibers, (26) secondary xylem cell matrix, (27) secondary growth pattern,
(28) axial parenchyma, (29) pith sclerenchyma, (30) pith collenchyma, (31) pith crystals, (32)
pith ducts, (33) pith wide-band tracheids, (34) pith mucilage, (35) leaf epidermal crystals, (36)
leaf epidermal mucilage, (37) leaf cuticle, (38) leaf epidermal projections, (39) leaf hypodermis,
(40) leaf hypodermal tannins, (41) photosynthetic type, (42) mesophyll ducts, (43) mesophyll
crystals, (44)mesophyll tannins, (45) mesophyll collenchyma, (46)mesophyll mucilage,
(47)midrib xylem cell majority, (48) midrib xylary fibers, (49) lateral xylem cell majority,
(50)stem succulence, (51)leaf succulence.
For phylogenetic analysis, Achatocarpus gracilis (Achatocarpaceae) was used as an
outgroup taxa to serve as a comparison to the succulent familial alliance of Aizoaceae,
Basellaceae, Cactaceae, Didiereaceae, and Portulaceae. Using PAUP*4.0, beta 8, parsimony
analysis was performed. A decay analysis was performed by using the heuristic search option
and keeping all trees between 150 and 160 steps.
RESULTS AND DISCUSSION
Of the 51 characters included in the parsimony analysis, 11 were found to be
parsimony-uninformative based on the complete data matrix, leaving 40 characters for analysis.
Heuristic searching produced one tree of 156 steps that was found on a single island. With the
result being unusual, the search was repeated 10 times with no change in topology.
Pereskia (Cactaceae) was allied with Achatocarpus (Achatocarpaceae); Talinella
(Portulaceae), Cistanthe (Portulaceae), Anredera (Basellaceae), Grahamia (Portulaceae), and
Talinopsis (Portulaceae) showed a similar relationship. Talinum (Portulaceae) was in a sister-
taxon position with the remaining 14 taxa. The genera sampled for Aizoaceae comprised one
terminal clade. Ceraria (Portulaceae) and Portulacaria (Portulaceae), Southern African
succulent tree genera, were found to be embedded with Alluaudia (Didiereaceae) and Didierea
(Didiereaceae). Anacampseros (Portulaceae) and Talinaria (Portulaceae) formed a sister group
alliance, and Portulaca (Portulaceae) was allied with Anredera (Basellaceae) and Tournonia
(Basellaceae). The decay analysis of trees longer than 156 steps indicated strong support, at
least in the 50% majority tree.
The resulting tree largely agrees with recent existing molecular phylogenies of this
group of succulent families. The most parsimonious tree produced shows that Pereskia
(Cactaceae) falls clearly into a Northern American grouping of genera, with the exception of the
Madagascan species Talinella pachypoda, supporting the general conclusion among recent
papers that Portulaceae is most likely paraphyletic. The grouping of genera from Aizoaceae is an
unexpected result of the analysis, due to the fact that the five sub-families were often segregated
into Aizoaceae (Aizoadeae, Sesuvioideae, and Tetragonioideae) and Mesembryanthemaceae
(Mesembryanthemonideae and Ruschioideae).
There is an overlap of biogeography and traits, especially in the arid regions of Southern
Africa, with more succulent genera of Aizoaceae and Portulaceae being mostly found as
endemics. Interestingly, genera of both families have evolved wide-band tracheid (WBTs) in leaf
xylem, as have subfamilies of Cactaceae. Two arguments arise from these findings: a
Gondwanaland homology or a case of convergent evolution.
First argument is that these tracheids are present due to ancestry, a common distant
ancestor, and are therefore a homologous evolutionary trait. However, since no published
phylogeny indicates a close alliance of Aizoaceae to either Cactaceae or Portulaceae, this
argument is not supported. The other argument would be that wide-band tracheid evolution is a
case of convergent evolution. The evidence of this lies in the degree of succulence of the plants
in which wide-band tracheids have evolved.
If this hypothesis were valid as well for Aizoaceae, then only genera with derived
succulent traits would have WBTs. This hypothesis has been demonstrated to be valid. The
hypothesis under investigation is that phylogeny is correlated with succulent trait evolution.
This can only be examined successfully in the context of paleoclimatic data and rates of
endemism.
Biogeographically, the four families are distinctly of a Gondwanaland distribution. The
separation of South America and Africa occurred between 80 and 90 million years ago. By that
time, Madagascar would have been far enough from Africa to prevent pollen exchange and seed
dispersal, resulting in reproductive isolation for ancestral Didiereaceae.
By the Miocene, deserts were forming in North America, South America, and Africa. By
the late Oligocene, a change in pre-existing rainfall patterns of Southern Africa from a mesic
regime to an aridiflying regime due to ocean current changes. Thus, by the Miocene, there were
definite signs of increasing aridification of Southern Africa and the isolation of Aizoaceae and
Portulaceae ancestral elements. By the Pliocene, South and North America became attached
allowing exchanges of flora and fauna.
An examination of isotope compositions of fossil soils and animal teeth estimated to be
about six million years old, and found a low level of carbon dioxide in the Miocene. A hypothesis
that this low level of carbon dioxide was the driving force for the modification of the

pathway
that leads to the evolution of the modern

pathway. Similar research found that low carbon

dioxide alone, without the accompanying influence of drier and warmer climatic conditions, was
insufficient to spur an increase in the distribution of

plants. Thus the

pathway probably
evolved in regimes with a combination of lower carbon dioxide levels and increasing
aridification.
The phylogenetic tree generated in this study and paleoclimatic data, in combination
with recent studies on the evolution of C4/CAM pathways, point to a relatively recent and rapid
evolution of succulent genera within these families. The simplest hypothesis id that (1)
stabilization of arid regions in the last five million years, and (2) accelerated evolution in species
that are reproductively isolated, have combined to create strong selective stress on the
ancestral populations, thus producing these unique succulents.

II. CRITIC
The journal of James V. Landrum entitled "Four Succulent Families and 40 Million Years
of Evolution and Adaptation to Xeric Environments: What Can Stem and Leaf Anatomical
Characters Tell Us About Their Phylogeny?" published by International Association for Plant
Taxonomy (IAPT) focuses on an evolutionary examination of stem and leaf anatomical
adaptations, and an attempt to link paleoclimatology to the evolution of these traits.
METHODOLOGY
Using only taxa with leaf and stem samples and getting the leaf transverse sections and
stem transverse, radial, and tangential sections taken at mid-leaf and mid-stem, respectively,
enable completion and consistency of data.
Navashin's solution was used to fixed living specimens and was embedded in Paraplast
Plus similar to what Mauseth, Montenegro & Walckowiak (1984) used in their journal. However,
instead of using a Navashin's solution Mauseth & al. used FAA solution. The characters used had
reliable bases like Schimper's (1903) xeromorphic characters and more modern interpretations
of succulent characters found in studies by Metcalfe & Chalke (1983), Carlquist (1988), Mauseth
& al. (1995), and Mauseth & Landrum (1997).
The author used the appropriate methods to gather the evidence by using the parsimony
and decay analysis. The differentiation of the outgroup sample or taxa with the ingroup taxa
made the methodology clearer. Parsimony analysis was performed using the PAUP*40, Beta 8
(Swofford, 1999) while the decay analysis was done by using Heuristic search options. The
methods used were modern and valid, especially the software, PAUP*40, Beta 8.
RESULTS AND DISCUSSION
Based on the results, 11 out of the 51 characters were found to be parsimony-
uninformative, leaving 40 characters for analysis. Stating that the 11 characters were
parsimony-uninformative or of no use in resolving the tree was able to help the readers
interpret the result of the analysis. By repeating the Heuristic search many times, the
production of that single most parsimonious tree became more reliable. Also, a strong support
was indicated by the decay analysis. The resulting tree supported with recent molecular
phylogenies of succulent families (Applequist & Wallace, 2000, 2001) cannot be questioned.
Recent and reliable papers of Downic & al. (1997) and Hershkovitz & Zimmer (1997) strongly
support the results obtained from the data.
The author presented two contrasting arguments which may be hypothesis for the
adaptation of succulent plants: the Gondwanaland homology and the case of convergent
evolution. The first view is not supported due to no or lack of evidence or published phylogeny.
The second argument is a valid hypothesis with strong evidence. The author showed two
contrasting ideas for the readers to think deeper and analyse the possibilities. The author did
not just did that to let the readers think deeper but also to accept the possibility of the first
argument to be valid once more evidences are gathered.
The second hypothesis, which is more valid compared to the first, used the context of
paleoclimatic data and rates of endemism as evidences. Valid, modern, and reliable papers by
Gentry (1982), Raven & Axelrod (1974), Tankard & Rogers (1978) and Gibson & Nobel (1986)
were used as reference and evidence to strengthen the second hypothesis.
The evolution of CAM pathways in succulent plants showed how these plants adapted to
xeric environments. The above statement can be supported by recent and valid researches of
Cerling & al. (1998), Nambudiri & al. (1978), Thomasson & al. (1986), and Ehleringer &
Monsoon (1998).
The author, with the used of valid, recent, modern and reliable sources and references,
stated the objective of the article, explained the methodology, results and discussion in a logical,
unbiased way and used the evidence to draw the reader to the same conclusion which made me
strongly agree with him.

III. CROSS-REFERENCE
A. Phylogeny of the Portulacaceous Cohort Based on ndhF Sequence Data
By Wendy L. Applequist and Robert S. Wallace
Abstract:
The Portulacaceae, Basellaceae, Cactaceae, and Didiereaceae form a monophyletic group
within the Caryophyllales, and evidence exists that the first may be paraphyletic if the latter
three are recognized at the familial level. Several taxonomic treatments of the Portulacaceae
based on morphological features have failed to produce consensus regarding infrafamilial
relationships. The present paper employs sequences of the chloroplast gene ndhF to produce a
molecular phylogeny of the portulacaceous cohort, focusing on the relationship among major
lineages of the Portulacaceae and the three families potentially derived from within them.
Results of this analysis supported two major clades within the monophyletic cohort. The first
included Portulaca, Anacampseros and its relatives, much of Talinum, Talinella, and Cactaceae;
the second, weakly supported, included the remaining genera of Portulacaceae, Basellaceae, and
Didiereaceae. This phylogeny also showed that several generic circumscriptions remain
inadequate, particularly that of Talinum sensu lato, which was polyphyletic in this analysis, and
that all present classifications of the Portulacaceae include demonstrably non-monophyletic
tribes.
Applequist and Wallaces journal, at first, appears to be quite similar to Landrums
journal; however, this is not so. The two journals may be similar in the taxa being
examined, Caryophyllales and both journals examines the phylogeny of the families
under Caryophyllales but only on that area. It should be noted that Applequist and
Wallaces journal was published first compared to Landrums journal. The journals are
different in such a way that the former examined the families Portulacaceae,
Basellaceae, Cactaceae, and Didiereaceae while the latter examined the families
Portulacaceae, Aizoaceae, Cactaceae, and Didiereaceae. They are also different in the
way the families were examined: the former using ndhF sequence data while the latter
using leaf and stem anatomical characters.
B. Phylogeny of the Madagascan endemic family Didiereaceae
By Wendy L. Applequist and Robert S. Wallace
Abstract:
A molecular phylogeny of the Didiereaceae was produced through parsimony analysis of
chloroplastrpl16 intron andtrnL-trnF andtrnT-trnL intergenic spacer sequences of all eleven
species of the Didiereaceae and several outgroup taxa from the Portulacaceae. Results indicated
that: 1) the Didiereaceae were embedded within the Portulacaceae, with Calyptrotheca as the
sister group of the family; 2) present generic limits were supported; 3) Alluaudiopsis was the
most basal lineage; 4) at least two separate episodes of polyploidization within the genus
Alluaudia had occurred, and 5) unusually low amounts of variation were present in rapidly
evolving noncoding plastid sequences.
Applequist and Wallaces journal is similar to Landrums journal in a way that both
journals study the family Didiereaceae of Caryophyllales. They differ in the area the
journals are covering. The former only focuses on family Didiereaceae while the latter
does not only focus on family Didiereaceae but also to Portulacaceae, Aizoaceae, and
Cactaceae. They also differ in the way the families were examined. In the former,
molecular characters were used while in the latter, anatomical characters were used.
C. Structural and systematic study of an unusual tracheid type in cacti
By James D. Mauseth, Yoriko Uozumi, Brandon J. Plemons, James V. Landrum
Abstract:
Wide-band tracheids are a specialized tracheid type in which an annular or helical
secondary wall projects deeply into the cell lumen. They are short, wide and spindle-shaped,
and their bandlike secondary walls cover little of the primary wall, leaving most of it available
for water diffusion. Wide-band tracheids appear to store and conduct water while preventing
the spread of embolisms. They may be the most abundant tracheary element in the xylem, but
they are always accompanied by at least a few vessels. Typically, fibers are absent wherever
wide-band tracheids are present. Wide-band tracheids occur in the primary and secondary
xylem of succulent stems, leaves and roots in genera of all three subfamilies of Cactaceae but
were not found in the relictual genus Pereskia, which lacks succulent tissues. In the large
subfamily Cactoideae, wide-band tracheids occur only in derived members, and wide-band
tracheids of North American Cactoideae are narrower and are aligned in a more orderly radial
pattern than those of South American Cactoideae. Wide-band tracheids probably arose at least
three times in Cactaceae.
Mauseth & al. journal is similar to Landrums journal in a way that they both deal with
families of Caryophyllales and both study wide-band tracheids. They differ in the area
the journals are covering. The former only focuses on family Cactaceae while the latter
does not only focuses on Cactaceae but also on Portulacaceae, Aizoaceae and
Didiereaceae. They also differ with the main objective. The former focuses on the
structure of WBTs while the latter focuses on the phylogeny and adaptation of the
succulent families of Caryophyllales.