Molecular and morphological analyses have elucidated phylogenetic relationships of two remarkable species in the Phallales: Aseroe floriformis and Phallus roseus. Genes from ATPase subunit 6 (atp6), the nuclear large subunit ribossomal DNA (nuc-LSU), and the second largest RNA polymerase II subunit (RPB2) underwent Bayesian and parsimony molecular analyses. Molecular datasets combined with morphological characters, support a new genus (Abrachium for Aseroe floriformis), reassessment Itajahya, and emendation of Clathraceae.
Inter Simple Sequence Repeats (ISSR) markers were utilized to identify the levels of heritable varieties and patterns of the populace structure among the five populaces of Pteris biaurita, a natural fern in India. A comprehensive examination was directed in three replicates at 2013-14 seasons in the Western Ghats, South India. Five wild P. biaurita, accessions (maiden hair) were assessed for genotyping studies. Results demonstrated a pivotal discrepancy among genotypes for they were characterized in view of this uniqueness in four groups by the genetic cluster examination. In this trial, ISSR primers amplified 63 polymorphic groups. In view of the genetic identity data, genotypes were figured and differed from 0.5714 to 0.6984. The percentage of polymorphism indicated predominant genotype that may be utilized for the conservation of species. ISSR appeared to be an obliging marker for prediction of genotype inside a closed group of inter specific populace in the investigation territory
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Inter Simple Sequence Repeats (ISSR) markers were utilized to identify the levels of heritable varieties and patterns of the populace structure among the five populaces of Pteris biaurita, a natural fern in India. A comprehensive examination was directed in three replicates at 2013-14 seasons in the Western Ghats, South India. Five wild P. biaurita, accessions (maiden hair) were assessed for genotyping studies. Results demonstrated a pivotal discrepancy among genotypes for they were characterized in view of this uniqueness in four groups by the genetic cluster examination. In this trial, ISSR primers amplified 63 polymorphic groups. In view of the genetic identity data, genotypes were figured and differed from 0.5714 to 0.6984. The percentage of polymorphism indicated predominant genotype that may be utilized for the conservation of species. ISSR appeared to be an obliging marker for prediction of genotype inside a closed group of inter specific populace in the investigation territory
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.
Colletotrichum causes anthracnose in crops around the world producing postharvest losses up to 60%. There are a great variety of Colletotrichum strains isolated from mango orchards. Thus, it is important to characterize their pathogenicity, as well as to perform a correct identification, in order to implement good strategies to eradicate the produced disease. The aim of this work is to identify Colletotrichum spp. and to determine the production of Pectate Lyase (PL) as a virulence factor in the pathogenicity process. Macroscopic characteristics of isolated colony vary from grey to salmon, sometimes showing luxuriant orange conidial masses with grey or white bottom. Conidia vary from 10.39 to 14.83 × 2.75 to 3.40 μm corresponding to C. gloeosporioides or C. acutatum according to Sutton. Growth rates vary from 0.1948 to 0.2239 day-1. The pectate lyase activity was induced by mango cells (240.81 VS 398U/L). According to CgInt and ITS4 PCR amplification M2V and SA correspond to C. gloeosporioides.
Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.
Colletotrichum causes anthracnose in crops around the world producing postharvest losses up to 60%. There are a great variety of Colletotrichum strains isolated from mango orchards. Thus, it is important to characterize their pathogenicity, as well as to perform a correct identification, in order to implement good strategies to eradicate the produced disease. The aim of this work is to identify Colletotrichum spp. and to determine the production of Pectate Lyase (PL) as a virulence factor in the pathogenicity process. Macroscopic characteristics of isolated colony vary from grey to salmon, sometimes showing luxuriant orange conidial masses with grey or white bottom. Conidia vary from 10.39 to 14.83 × 2.75 to 3.40 μm corresponding to C. gloeosporioides or C. acutatum according to Sutton. Growth rates vary from 0.1948 to 0.2239 day-1. The pectate lyase activity was induced by mango cells (240.81 VS 398U/L). According to CgInt and ITS4 PCR amplification M2V and SA correspond to C. gloeosporioides.
ARTICLE IN PRESShttpwww.elsevier.deprotisPublished onl.docxfredharris32
ARTICLE IN PRESS
http://www.elsevier.de/protis
Published online date 7 August 2006
1
Correspondin
fax +81 29 853
e-mail iinouye
2Current addr
Parkville, Victo
& 2006 Elsev
doi:10.1016/j
157, 401—419, August 2006
Protist, Vol.
ORIGINAL PAPER
Hatena arenicola gen. et sp. nov., a Katablepharid
Undergoing Probable Plastid Acquisition
Noriko Okamoto2, and Isao Inouye1
Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba,
Ibaraki 305-8572, Japan
Submitted February 27, 2006; Accepted May 27, 2006
Monitoring Editor: Robert A. Andersen
Hatena arenicola gen. et sp. nov., an enigmatic flagellate of the katablepharids, is described. It shows
ultrastructural affinities to the katablepharids, including large and small ejectisomes, cell covering,
and a feeding apparatus. Although molecular phylogenies of the 18S ribosomal DNA support its
classification into the katablepharids, the cell is characterized by a dorsiventrally compressed cell
shape and a crawling motion, both of which are unusual within this group. The most distinctive feature
of Hatena arenicola is that it harbors a Nephroselmis symbiont. This symbiosis is distinct from
previously reported cases of ongoing symbiosis in that the symbiont plastid is selectively enlarged,
while other structures such as the mitochondria, Golgi body, cytoskeleton, and endomembrane
system are degraded; the host and symbiont have developed a morphological association, i.e., the
eyespot of the symbiont is always at the cell apex of Hatena arenicola; and only one daughter cell
inherits the symbiont during cell division, resulting in a symbiont-bearing green cell and a symbiont-
lacking colorless cell. Interestingly, the colorless cells have a feeding apparatus that corresponds to
the location of the eyespot in symbiont-bearing cells, and they are able to feed on prey cells. This
indicates that the morphology of the host depends on the presence or absence of the symbiont. These
observations suggest that Hatena arenicola has a unique ‘‘half-plant, half-predator’’ life cycle; one cell
divides into an autotrophic cell possessing a symbiotic Nephroselmis species, and a symbiont-lacking
colorless cell, which later develops a feeding apparatus de novo. The evolutionary implications of
Hatena arenicola as an intermediate step in plastid acquisition are discussed in the context of other
examples of ongoing endosymbioses in dinoflagellates.
& 2006 Elsevier GmbH. All rights reserved.
Key words: Hatena arenicola; Katablepharidophyta/Kathablepharida; Nephroselmis symbiont; plant
evolution; plastid acquisition via secondary endosymbiosis; ultrastructure.
Abbreviations: EM ¼ electron microscopy; ER ¼
endoplasmic reticulum; ICBN ¼ International Code
of Botanical Nomenclature; ICZN ¼ International
Code of Zoological Nomenclature; LM ¼ light mi-
croscopy; SEM ¼ scanning electron microscopy;
SSU rDNA ¼ small subunit ribosomal DNA; TEM ¼
transmission electron microscopy.
g author;
4533
...
Microsatellite and mt-DNA phylogenies of the chamois (genus Rupicapra) and ta...Trinidad Mendez
To elucidate the evolutionary history of chamois, we had analysed DNA sequences of four mitochondrial regions and 20 loci microsatellites including all subspecies along its entire distribution range
Climate Impact of Software Testing at Nordic Testing DaysKari Kakkonen
My slides at Nordic Testing Days 6.6.2024
Climate impact / sustainability of software testing discussed on the talk. ICT and testing must carry their part of global responsibility to help with the climat warming. We can minimize the carbon footprint but we can also have a carbon handprint, a positive impact on the climate. Quality characteristics can be added with sustainability, and then measured continuously. Test environments can be used less, and in smaller scale and on demand. Test techniques can be used in optimizing or minimizing number of tests. Test automation can be used to speed up testing.
Communications Mining Series - Zero to Hero - Session 1DianaGray10
This session provides introduction to UiPath Communication Mining, importance and platform overview. You will acquire a good understand of the phases in Communication Mining as we go over the platform with you. Topics covered:
• Communication Mining Overview
• Why is it important?
• How can it help today’s business and the benefits
• Phases in Communication Mining
• Demo on Platform overview
• Q/A
Threats to mobile devices are more prevalent and increasing in scope and complexity. Users of mobile devices desire to take full advantage of the features
available on those devices, but many of the features provide convenience and capability but sacrifice security. This best practices guide outlines steps the users can take to better protect personal devices and information.
UiPath Test Automation using UiPath Test Suite series, part 5DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 5. In this session, we will cover CI/CD with devops.
Topics covered:
CI/CD with in UiPath
End-to-end overview of CI/CD pipeline with Azure devops
Speaker:
Lyndsey Byblow, Test Suite Sales Engineer @ UiPath, Inc.
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
How to Get CNIC Information System with Paksim Ga.pptxdanishmna97
Pakdata Cf is a groundbreaking system designed to streamline and facilitate access to CNIC information. This innovative platform leverages advanced technology to provide users with efficient and secure access to their CNIC details.
A tale of scale & speed: How the US Navy is enabling software delivery from l...sonjaschweigert1
Rapid and secure feature delivery is a goal across every application team and every branch of the DoD. The Navy’s DevSecOps platform, Party Barge, has achieved:
- Reduction in onboarding time from 5 weeks to 1 day
- Improved developer experience and productivity through actionable findings and reduction of false positives
- Maintenance of superior security standards and inherent policy enforcement with Authorization to Operate (ATO)
Development teams can ship efficiently and ensure applications are cyber ready for Navy Authorizing Officials (AOs). In this webinar, Sigma Defense and Anchore will give attendees a look behind the scenes and demo secure pipeline automation and security artifacts that speed up application ATO and time to production.
We will cover:
- How to remove silos in DevSecOps
- How to build efficient development pipeline roles and component templates
- How to deliver security artifacts that matter for ATO’s (SBOMs, vulnerability reports, and policy evidence)
- How to streamline operations with automated policy checks on container images
Generative AI Deep Dive: Advancing from Proof of Concept to ProductionAggregage
Join Maher Hanafi, VP of Engineering at Betterworks, in this new session where he'll share a practical framework to transform Gen AI prototypes into impactful products! He'll delve into the complexities of data collection and management, model selection and optimization, and ensuring security, scalability, and responsible use.
Essentials of Automations: The Art of Triggers and Actions in FMESafe Software
In this second installment of our Essentials of Automations webinar series, we’ll explore the landscape of triggers and actions, guiding you through the nuances of authoring and adapting workspaces for seamless automations. Gain an understanding of the full spectrum of triggers and actions available in FME, empowering you to enhance your workspaces for efficient automation.
We’ll kick things off by showcasing the most commonly used event-based triggers, introducing you to various automation workflows like manual triggers, schedules, directory watchers, and more. Plus, see how these elements play out in real scenarios.
Whether you’re tweaking your current setup or building from the ground up, this session will arm you with the tools and insights needed to transform your FME usage into a powerhouse of productivity. Join us to discover effective strategies that simplify complex processes, enhancing your productivity and transforming your data management practices with FME. Let’s turn complexity into clarity and make your workspaces work wonders!
zkStudyClub - Reef: Fast Succinct Non-Interactive Zero-Knowledge Regex ProofsAlex Pruden
This paper presents Reef, a system for generating publicly verifiable succinct non-interactive zero-knowledge proofs that a committed document matches or does not match a regular expression. We describe applications such as proving the strength of passwords, the provenance of email despite redactions, the validity of oblivious DNS queries, and the existence of mutations in DNA. Reef supports the Perl Compatible Regular Expression syntax, including wildcards, alternation, ranges, capture groups, Kleene star, negations, and lookarounds. Reef introduces a new type of automata, Skipping Alternating Finite Automata (SAFA), that skips irrelevant parts of a document when producing proofs without undermining soundness, and instantiates SAFA with a lookup argument. Our experimental evaluation confirms that Reef can generate proofs for documents with 32M characters; the proofs are small and cheap to verify (under a second).
Paper: https://eprint.iacr.org/2023/1886
GraphSummit Singapore | The Future of Agility: Supercharging Digital Transfor...Neo4j
Leonard Jayamohan, Partner & Generative AI Lead, Deloitte
This keynote will reveal how Deloitte leverages Neo4j’s graph power for groundbreaking digital twin solutions, achieving a staggering 100x performance boost. Discover the essential role knowledge graphs play in successful generative AI implementations. Plus, get an exclusive look at an innovative Neo4j + Generative AI solution Deloitte is developing in-house.
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Today, after several years of existence, an extremely active community and an ultra-dynamic ecosystem, Kubernetes has established itself as the de facto standard in container orchestration. Thanks to a wide range of managed services, it has never been so easy to set up a ready-to-use Kubernetes cluster.
However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
In his public lecture, Christian Timmerer provides insights into the fascinating history of video streaming, starting from its humble beginnings before YouTube to the groundbreaking technologies that now dominate platforms like Netflix and ORF ON. Timmerer also presents provocative contributions of his own that have significantly influenced the industry. He concludes by looking at future challenges and invites the audience to join in a discussion.
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Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
Removing Uninteresting Bytes in Software FuzzingAftab Hussain
Imagine a world where software fuzzing, the process of mutating bytes in test seeds to uncover hidden and erroneous program behaviors, becomes faster and more effective. A lot depends on the initial seeds, which can significantly dictate the trajectory of a fuzzing campaign, particularly in terms of how long it takes to uncover interesting behaviour in your code. We introduce DIAR, a technique designed to speedup fuzzing campaigns by pinpointing and eliminating those uninteresting bytes in the seeds. Picture this: instead of wasting valuable resources on meaningless mutations in large, bloated seeds, DIAR removes the unnecessary bytes, streamlining the entire process.
In this work, we equipped AFL, a popular fuzzer, with DIAR and examined two critical Linux libraries -- Libxml's xmllint, a tool for parsing xml documents, and Binutil's readelf, an essential debugging and security analysis command-line tool used to display detailed information about ELF (Executable and Linkable Format). Our preliminary results show that AFL+DIAR does not only discover new paths more quickly but also achieves higher coverage overall. This work thus showcases how starting with lean and optimized seeds can lead to faster, more comprehensive fuzzing campaigns -- and DIAR helps you find such seeds.
- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
2. 420 ... Cabral & al.
the presence of a campanula (head) at the apex of the receptacle in Phallus,
indusium in Dictyophora, and calyptra in Itajahya. Some genera lack a distinct
head and form the mucilaginous gleba directly on the apical receptacle (e.g.,
Mutinus) or at a constriction near the apex (e.g., Staheliomyces). Several
attempts were made to establish a synopsis of the family (Lloyd 1909, Fischer
1933), and Cunningham (1944) included seven genera: Dictyophora, Itajahya,
Phallus, Mutinus, Staheliomyces, Aporophallus, and Floccomutinus. However,
Kreisel (1996) viewed Dictyophora, Aporophallus, Itajahya, Echinophallus, and
Endophallus as subgenera of Phallus sensu lato, using the shape and surface
of the head, pigmentation of the receptacle, and morphology of the volva and
rhizomorphs as primary characteristics. He did not deem the presence of a pore
at the receptacle apex or the morphology of the head, indusium, and calyptra as
sufficient for segregation at the generic level (Kreisel 1996).
Taxonomic studies of Neotropical phalloid species have revealed new
and intriguing species (Baseia et al. 2003, Baseia & Calonge 2005, Fazolino
et al. 2010). The discovery of new species is essential to understanding the
evolutionary path of Phallales clades, since according to known estimates
(Hawksworth 2001, Mueller et al. 2007, Blackwell 2011), most tropical species
remain unknown. Through morphological data and molecular tools the
classification of macrofungi should change in coming years.
The present study aims to evaluate the actual taxonomic position of Aseroe
floriformis and Phallus roseus, two species with distinct morphological features
that have not yet been analyzed by molecular studies.
Materials & methods
Basidioma sampling
Basidiomata were sampled at the Parque Estadual Dunas do Natal (06°22ʹ10ʺ S
and 35°00ʹ28ʺ W) and Estação Ecológica do Seridó (06°35ʹ02ʺ S and 37°17ʹ02ʺ W),
Rio Grande do Norte state, Brazil. Specimens of Aseroe floriformis and Phallus roseus
were photographed and collected during the rainy seasons in 2008 and 2009. Materials
studied were deposited in the fungal collection of the UFRN Herbarium.
Macroscopic characters were described from fresh specimens. Basidiomata colors
were based on Kornerup & Wanscher (1978). Microscopic characteristics were
determined from temporary mounts, hand-sectioned with a razor blade on the vertical
axis of the basidiomata and mounted in 3% KOH and 1% cotton-blue lactophenol,
respectively. All measurements were made under oil-immersion microscope objective
at 1000× magnification. Spore measurements included the largest and smallest spores
and at least 20 additional randomly selected spores from each specimen.
Molecular techniques
Fresh basidiomata of Aseroe floriformis (UFRN 1481) and Phallus roseus (UFRN
535) were used for CTAB DNA extraction. The CTAB buffer (1.4M NaCl, 100mM Tris-
HCl pH 8.20mM EDTA and 2% CTAB) was added to 0.8g of fresh tissue pulverized
3. Abrachium floriforme gen. & sp. nov. (Brazil) ... 421
with liquid nitrogen and incubated for 30 min. at 65°C. After incubation, a chloroform-
isoamyl (24:1) extraction was performed, followed by isopropanol precipitation. Total
DNA amount was quantified on a NanoDrop-2000 spectrophotometer (NanoDrop
Technologies Inc. Wilmington, DE 1980, United States).
DNA sequences were obtained for three different regions: nuc-LSU-rDNA (LROR/
LR5), ATPase subunit 6 (ATP61/ATP62) and the second largest subunit of RNA
polymerase RPB2 (bRPB2-6F/bRPB2-7R), with previously designed primers (Vilgalys
1990; Liu et al. 1999; Kretzer & Bruns 1999).
All PCR reactions were carried out in a final volume of 50µl, with PCR buffer 10X,
4 mM of MgCl2, 25 ng of DNA, 0.3 mM of dNTP Mix, 12.5 pmol of each primer and
0.3µl of Invitrogen® DNA Taq Polymerase (5U/µl). The initial cycling parameter for
atp6 reaction condition consisted of 2 min at 95°C for initial denaturation, 5 cycles of 35
s at 94°C, 55 s at 37°C and 1min at 72°C. This was followed by 30 cycles of 35 s at 94°C,
55 s at 45°C, 1 min at 72°C, and final extension of 5 min at 72°C with final hold of 4°C.
The nuc-LSU-rDNA amplification condition underwent initial denaturation for 2 min
at 95°C, followed by 39 cycles for 1 min at 94°C, 30 s annealing at 45°C, 1 min extension
at 72°C, and a final extension for 5min at 72°C. RPB2 amplifications followed conditions
described by Liu et al. (1999).
Amplicons were verified by ethidium bromide-stained 0.4% agarose gel
electrophoresis. Total amount of amplified DNA was quantified by a NanoDrop-
2000 spectrophotometer. Amplifications with low DNA content were re-amplified by
increasing PCR cycles.
PCR products were purified with Wizard® SV Gel and PCR Clean-Up System
(Promega). The sequencing reaction was performed with a DNA sequencing kit
(BigDye® Terminator v3.1 Cycle Sequencing Kit) using the same primers employed to
amplify each fragment, and sequencing was conducted an ABI 3730 DNA Analyzer
(Applied Biosystems). Both were carried out at the Center for Human Genome Studies
(University of São Paulo, CEGH-USP) and later assembled using DNA Baser v3.0.12
beta.
Phylogenetic analysis
Phylogenetic analyses were performed for three combined DNA datasets under
Bayesian and parsimony criteria. Newly generated sequences for Aseroe floriformis and
Phallus roseus ATP6, nuc-LSU-rDNA and RPB2 and those retrieved from GenBank
(National Center for Biotechnology Information, http://www.ncbi.nlm.nih.gov – Table
1) were separately aligned using ClustalX2 (Larkin et al. 2007), with default settings
(alignment files are available at http://purl.org/phylo/treebase/phylows/study/TB2:
S11546). Manual adjustments were made in BioEdit v. 7.0.9.0 (Hall 1999). The three
alignments were then concatenated with Phyutility (Smith & Dunn 2008). Maximum
parsimony analyses were conducted with combined dataset file, in PAUP* (Phylogenetic
Analysis Using Parsimony) version 4b10 (Swofford, 2002). Heuristic search was used with
tree bisection-reconnection (TBR) and MULTrees. Confidence levels were established
through 1000 bootstrap replicates. Bayesian analysis was carried out with combined
dataset in MrBayes v.3.1.2 (Huelsenbeck & Ronquist 2001). Two different runs with
four incrementally heated simultaneous Monte Carlo Markov chains were conducted
over 2 million generations, applying respective substitution models determined for
4. 422 ... Cabral & al.
each partition in MrModeltest v.2.2 (http://www.abc.se/~nylander/). Trees were
sampled every 100 generations to produce 20,000 trees. In order to estimate posterior
probabilities, 25% of the trees were discarded as a burn-in stage, observing when average
standard deviation of split frequency (ASDSF) values dropped below 0.01.
Table 1. GenBank accession numbers used for phylogenetic analysis.
GenBank Accession Number
Species
nucLSU ATP6 RPB2
Anthurus archeri DQ218624 DQ218913 DQ219081
Abrachium floriforme JF968440 JF968438 JF968442
Aseroe rubra DQ218625 DQ218914 DQ219082
Clathrus chrysomycelinus DQ218626 DQ218915 DQ219083
Claustula fischeri K.M. Curtis — — DQ219086
Dictyophora duplicata (Bosc) E. Fisch. DQ218481 DQ218765 DQ219087
D. indusiata (Vent.) Desv. DQ218627 DQ218917 DQ219088
D. multicolor Berk. & Broome DQ218628 DQ218918 DQ219089
Gelopellis sp. DQ218630 DQ218919 DQ219090
Gelopellis sp. DQ218631 DQ218920 DQ219091
Ileodictyon cibarium DQ218633 DQ218922 DQ219093
I. gracile DQ218636 DQ218925 DQ219096
Itajahya rosea JF968441 JF968439 —
Kobayasia nipponica (Kobayasi)
DQ218638 DQ218926 DQ219098
S. Imai & A. Kawam.
Laternea triscapa DQ218640 DQ218928 DQ219099
Lysurus borealis (Burt) Henn. DQ218641 DQ218929 DQ219100
L. mokusin (L.) Fr. DQ218507 DQ218791 DQ219101
Mutinus elegans (Mont.) E. Fisch. AY574643 AY574785 DQ219102
*Phallobata alba G. Cunn. DQ218642 DQ218930 DQ219103
*Phallus costatus (Penz.) Lloyd DQ218513 DQ218797 DQ219104
P. hadriani Vent. DQ218514 DQ218798 DQ219044
P. ravenelii Berk. & M.A. Curtis DQ218515 DQ218799 DQ219105
Protubera borealis S. Imai DQ218516 DQ218800 DQ219106
P. canescens DQ218645 DQ218932 DQ219108
P. jamaicensis (Murrill) Zeller DQ218647 DQ218933 DQ219110
P. maracuja Möller DQ218518 DQ218802 DQ219111
P. parvispora Castellano & Beever DQ218648 DQ218934 DQ219112
P. sabulonensis Malloch DQ218649 DQ218935 DQ219113
Simblum sphaerocephalum Schltdl. DQ218521 DQ218806 DQ219115
*Trappea darkeri (Zeller) Castellano DQ218651 DQ218938 DQ219116
Pseudocolus fusiformis AF213128 — —
* Species used as outgroup
5. Abrachium floriforme gen. & sp. nov. (Brazil) ... 423
Results
Phylogenetics analysis
Bayesian and maximum parsimony analyses produced no significant
differences in tree topology, indicating data consistency to resolve the
phylogeny of the group. The phylogram (Fig. 3) illustrates six well-formed
clades corresponding to the families of Phallales with high support values
(PP = 1, BT = 90–100). Intergeneric relationships in each family are also
shown. It was found that Aseroe floriformis does not cluster with Aseroe rubra,
and occupies a basal position in the Clathraceae, with PP = 1 and BT = 99.
In Phallaceae, Phallus roseus occupies a position between Mutinus elegans and
other Phallus species, including indusiate representatives (Dictyophora), which
cluster into a single clade displaying high levels of reliability (PP = 1, BT = 94).
Taxonomic revision
Based on the morphological and molecular data set, we propose a new genus
to accommodate Aseroe floriformis. The most important morphological feature
of the new genus —the lack of arms— requires an emendation in Clathraceae.
Phylogenetic analyses also support the status change of the genus Itajahya in
Phallaceae.
Clathraceae Chevall. emend. Baseia
Emended description: Basidiomata sessile to shortly stalked, upright
receptacle, spreading, sometimes arched and united arms or a latticed
receptaculum composed of interconnected tubular or gelatinous arms
expanding from a flaccid button, which acts as a volva, or armless sunflower-
shaped receptacle. Gleba located on the central disc of receptacle or inside
and next to the clathrate receptaculum. Peridium three-layered. Basidiospores
narrowly elliptical, covered with mucilaginous layer.
Type genus: Clathrus P. Micheli ex L.
Other genera: Abrachium Baseia & T.S. Cabral, Aseroe Labill.,
Blumenavia Möller, Pseudocolus Lloyd, Lysurus Fr., Ileodictyon Tul. & C.
Tul.
The genus Aseroe was proposed by Labillardière (1800), with Aseroe rubra
Labill. as the type (and only) species. Fischer (1890) subsequently described
A. arachnoidea. Yoshimi & Hongo (1989) added A. coccinea, a name recently
validated by Kasuya (2007). Baseia & Calonge (2005) discovered a new species,
Aseroe floriformis, in northeastern Brazil in sand dunes. Although Index
Fungorum lists 27 names in Aseroe, the latest edition of the Dictionary of
Fungi (Kirk et al. 2008) lists only two: A. floriformis and A. rubra. The genus
has a pantropical distribution, although A. rubra has been reported in Europe
(Spooner 2005). Aseroe is characterized by a cylindrical receptacle and stipe
6. 424 ... Cabral & al.
that emerge from a whitish volva. With the exception of Aseroe floriformis, all
species have apical free arms internally covered by the gleba (Dring 1980).
Abrachium Baseia & T.S. Cabral, gen. nov.
MycoBank MB 561618
Ovo subglobosae, epigaeo, rhizomorphis basalis. Stipite cylindrico, spongioso. Receptaculum
floriformis cum disco sine ramulis, centralis perforatae cum gleba gelatinosa marginata,
foetida. Sporis cylindraceo-bacilliformis, laevis, hyalinae.
Type species: Abrachium floriforme (Baseia & Calonge) Baseia & T.S. Cabral
Etymology: Abrachium, due to the absence of arms.
Key characters: Egg subglobose, epigeous with several and basal mycelial
cords, white. Stipe cylindrical, spongy. Receptacle with the same anatomy,
sunflower-shaped, without arms, central disc covered by a gelatinous gleba.
Spores cylindrical to bacilloid, smooth, hyaline.
Abrachium floriforme (Baseia & Calonge) Baseia & T.S. Cabral, comb. nov. Fig 1
MycoBank MB 561619
≡ Aseroe floriformis Baseia & Calonge, Mycotaxon 92: 170, 2005.
Specimens examined: BRAZIL, Rio Grande do Norte, Natal, Parque das Dunas,
ad solum arenarius, 3-VII-2004, legit I.G. Baseia & P.P.T. Lacerda, UFRN-Fungi 1851
(holotype); 07-V-2009, legit I.G. Baseia UFRN 1481, GenBank JF968440, JF968438,
JF968442.
Our phylogenetic analyses confirm high support values for Phallales in
accordance with Hosaka et al. (2007). Morphological and molecular analyses
supported the establishment of the genus Abrachium, named for its armless
receptacle (Fig 1). This taxon was originally described in the Brazilian Atlantic
rainforest (Baseia & Calonge 2005), but has since been recorded in northeastern
Brazil (Trierveiler-Pereira & Baseia 2009, 2011, Bezerra et al. 2009).
The clathraceous clade comprises members with a variable morphology of
the basidiomata, consisting of tubular or gelatinous arms either interconnected
or free (Dring 1980). The armless receptacle of Abrachium justifies an
emendation in Clathraceae and seems to be a primitive characteristic of the
clade, as shown in the tree obtained by molecular inference (Fig. 3). Molecular
data indicate that receptacle morphology is a phylogenetically informative
characteristic in Clathraceae. In phylogram, Abrachium represents a basal
clade, which is a sister group of a clade formed by other Clathraceae species.
The latter have a bifurcation leading to two other subgroups. The first subgroup
either has arms attached at the ends [Pseudocolus fusiformis (E. Fisch.) Lloyd],
or free and arched like a star [Aseroe rubra and Anthurus archeri (Berk.) E.
Fisch.]. The second subgroup displays a complex structure, forming a “cage”
type configuration including Clathrus chrysomycelinus Möller, Laternea
triscapa Turpin, Ileodictyon cibarium Tul. & C. Tul., Ileodictyon gracile Berk.,
7. Abrachium floriforme gen. & sp. nov. (Brazil) ... 425
Figures 1–2. Fresh specimens: 1. Abrachium floriforme. 2. Itajahya rosea. Bar = 10 mm.
and Protubera canescens G. W. Beaton & Malajczuk. Based on these data,
we can infer that the evolution to receptacle bifurcation (forming arms, e.g.,
Aseroe) occurred at least once in the Neotropical region. Presumably, after the
divergence of A. floriforme, a common ancestor has diverged to forms where
the receptacle may bifurcate like arms in one group, and form a cage in the
other (such as Ileodictyon).
Itajahya rosea (Delile) E. Fisch., Ber. Deutsch. Bot. Ges. 47: 294, 1929. Fig. 2
≡ Phallus roseus Delile, Flore d’Égypte: 300, 1813.
Specimen examined: BRAZIL, Rio Grande do Norte, Serra Negra do Norte,
Estação Ecológica do Seridó, ad solum arenarius, 23-V-2008, legit T.B.S. Ottoni, UFRN-
Fungi 535; GenBank JF968441, JF968439.
Molecular data analysis of Phallus roseus points to the need for reassessing
its current taxonomic status in the subgenus Itajahya proposed by Malençon
(1984) and Kreisel (1996). Itajahya was established by Möller (1895) based on I.
galericulata Möller, a taxon discovered in Santa Catarina state, Brazil. The main
feature distinguishing Itajahya from other Phallaceae genera is the presence
of a structure called a calyptra (Fig. 2) located at the apex of the gleba (Möller
1895, Malençon 1953, Ottoni et al. 2010). Molecular data demonstrated that
P. roseus diverged after Mutinus and prior to other species of Phallus (Fig.
3). Since I. rosea does not cluster with any other Phallus species, we propose
8. 426 ... Cabral & al.
Figure 3. Phylogenetic tree of Phallales obtained through Bayesian analysis of atp6, nuc-LSU
rRNA, and RPB2 sequences. Numbers preceding ‘/’ = Bayesian posterior probability values (PP)
and after ‘/’ = maximum parsimony bootstrap values (BT).
9. Abrachium floriforme gen. & sp. nov. (Brazil) ... 427
raising Itajahya to generic status. Further collections and molecular studies of
I. galericulata (= Phallus galericulatus (Möller) Kreisel) are also needed in order
to ensure morphological characteristics (mainly the presence of calyptra) agree
with molecular data recognizing the genus Itajahya.
Molecular data also imply that the indusium is not a phylogenetically
informative character in Phallaceae. Malençon (1984) and Kreisel (1996)
stated that species may be closely related with or without this structure, and
that a species may or may not produce it or have a rudimentary form of it.
The phylogram indicates that although Dictyophora multicolor and D. duplicata
form a separate clade, D. indusiata clusters with Phallus hadriani, exhibiting
prior divergence with the other mentioned clade. This suggests that indusiate
species may have emerged independently during several phases of the group’s
evolution, thereby characterizing it as a polyphyletic assemblage. The phylogram
also illustrates that Mutinus is a more primitive genus in Phallaceae, as observed
by Cunningham (1944), based on data from simple sporophore morphology
and the absence of complex structures such as campanulate apex, indusium
and calyptra. It is important to note that further molecular studies are needed,
especially with key-taxa that may lead to a change in current classification and
give us a clearer picture of the systematics and evolution of phalloid fungi.
Acknowledgments
We thank CNPq for the Msc. Scholarship and PPBio Semi-Arid Project for
financial support for this study. Sincere thanks are given to Prof José Luiz Bezerra
(Universidade Estadual de Santa Cruz) for encouraging this work, to Prof Vagner
Cortez for presubmission review, and to Prof. Gladstone Alves da Silva (Departamento
de Micologia, Universidade Federal de Pernambuco) for critical comments of molecular
analyses.
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